Information on EC 3.6.5.3 - protein-synthesizing GTPase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea

EC NUMBER
COMMENTARY
3.6.5.3
-
RECOMMENDED NAME
GeneOntology No.
protein-synthesizing GTPase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
GTP + H2O = GDP + phosphate
show the reaction diagram
this enzyme comprises a family of proteins involved in prokaryotic as well as eukaryotic protein synthesis. In the initiation factor complex, it is IF-2b (98kDa) that binds GTP and subsequently hydrolyses it in prokaryotes. in eukaryotes, it is eIF-2 (150 kDa) that binds GTP. In the elongation phase, the GTP-hydrolysing proteins are the EF-Tu polypeptide of the prokaryotic transfer factor (43 kDa), the eukaryotic elongation factor EF-1a (53 kDa), the prokaryotic EF-G (77 kDa), the eukaryotic EF-2 (70-110 kDa) and the signal recognition particle that play a role in endoplasmic reticulum protein synthesis (325 kDa). EF-Tu and EF-1a catalyse binding of aminoacyl-tRNA to the ribosomal A-site, while EF-G and EF-2 catalyse the translocation of peptidyl-tRNA from the A-side to the P-side. GTPase activity is also involved in polypeptide release from the ribosome with the aid of the pRFs and eRFs
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis
Q01698
-
hydrolysis
Saccharomyces cerevisiae NOY891, Thermus thermophilus JC469
-
-
-
hydrolysis of phosphoric ester
-
-
-
-
hydrolysis of phosphoric ester
-
-
additional information
-
influence on translation initiation pathway and ribosomal subunit joining
additional information
-
anti-association activity for splitted 70S ribosomes subunits; together with ribosome recycling factor and GTP transient split of 70S ribosomes into subunits
additional information
Escherichia coli DH5alpha
-
anti-association activity for splitted 70S ribosomes subunits; together with ribosome recycling factor and GTP transient split of 70S ribosomes into subunits
-
PATHWAY
KEGG Link
MetaCyc Link
NIL
-
SYSTEMATIC NAME
IUBMB Comments
GTP phosphohydrolase (mRNA-translation-assisting)
This enzyme comprises a family of proteins involved in prokaryotic as well as eukaryotic protein synthesis. In the initiation factor complex, it is IF-2b (98 kDa) that binds GTP and subsequently hydrolyses it in prokaryotes. In eukaryotes, it is eIF-2 (150 kDa) that binds GTP. In the elongation phase, the GTP-hydrolysing proteins are the EF-Tu polypeptide of the prokaryotic transfer factor (43 kDa), the eukaryotic elongation factor EF-1alpha (53 kDa), the prokaryotic EF-G (77 kDa), the eukaryotic EF-2 (70-110 kDa) and the signal recognition particle that play a role in endoplasmic reticulum protein synthesis (325 kDa). EF-Tu and EF-1alpha catalyse binding of aminoacyl-tRNA to the ribosomal A-site, while EF-G and EF-2 catalyse the translocation of peptidyl-tRNA from the A-site to the P-site. GTPase activity is also involved in polypeptide release from the ribosome with the aid of the pRFs and eRFs.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
aEF-1alpha
Sulfolobus solfataricus MT-4
-
-
-
archaeal initiation factor 2
-
-
archaeal initiation factor 2
Q980A5
-
archaeal initiation factor 2C
-
-
archaeal translation initiation factor 2
Q980A5
-
chloroplast elongation factor G
-
-
EC 3.6.1.48
-
-
formerly
-
Eco-IF2
-
-
eEF1A
P13905
genes A1, A2, A3, A4
EF-1alpha
P35021
-
EF-G
Escherichia coli DH5alpha, Escherichia coli MRE600
-
-
-
EF-G GTPase
-
-
EF-G1mt
-
-
EF-like GTPase
-
a class of eukaryotic GTPase with a punctate distribution suggesting multiple functional replacements of translation elongation factor 1alpha
EF-Tu
P0CE48
-
EF-Tu
Q01698
-
EF-Tu
P60338
-
EF-Tu
Thermus thermophilus JC469
-
-
-
EF-Tumt
-
-
EF4
Escherichia coli MRE600
-
-
-
eIEF2
-
-
eIF2
Saccharomyces cerevisiae J293
-
-
-
eIF2alpha
-
-
eIF5B
Saccharomyces cerevisiae NOY891
P39730
-
-
elongation factor
-
-
elongation factor (EF)
-
-
-
-
elongation factor 1 alpha
-
-
elongation factor 1 alpha
Sulfolobus solfataricus MT-4
-
-
-
elongation factor 1alpha
P35021
-
elongation factor 2
-
-
elongation factor 4
-
-
elongation factor 4
Escherichia coli MRE600
-
-
-
elongation factor G
Escherichia coli DH5alpha, Escherichia coli MRE600
-
-
-
elongation factor G
-
-
elongation factor Tu
P0CE48
-
elongation factor Tu
-
-
elongation factor Tu
Q01698
-
elongation factor Tu
-
-
elongation factor Tu
P60338
-
elongation factor Tu
Thermus thermophilus JC469
-
-
-
eukaryotic elongation factor one alpha
P13905
-
eukaryotic initiation factor 2
-
-
eukaryotic initiation factor 2A
-
-
eukaryotic initiation factor 5B
-
-
eukaryotic translation initiation factor 2
-
-
GTP phosphohydrolase
-
-
-
-
GTPase
-
-
-
-
GTPase HflX
Q980M3
-
GTPase HflX
Q980M3
-
-
GTPase-activating protein
-
-
guanine triphosphatase
-
-
-
-
guanine-nucleotide-exchange factor of eIF2
P32501
-
guanosine 5'-triphosphatase
-
-
-
-
guanosine triphosphatase
-
-
-
-
HflX GTPase
Q980M3
-
HflX GTPase
Q980M3
-
-
IF3
Escherichia coli DH5alpha
-
-
-
initiation factor (IF)
-
-
-
-
initiation factor 2
-
-
initiation factor 2
-
-
initiation factor 3
-
-
initiation factor 3
Escherichia coli DH5alpha
-
-
-
initiation factor-2
-
-
LepA
Escherichia coli MRE600
-
-
-
mitochondrial elongation factor G
-
-
mitochondrial initiation factor 2
-
-
peptide-release or termination factor
-
-
-
-
protein-synthesizing GTPase
-
-
protein-sythesizing GTPase
-
-
protein-sythesizing GTPase
P35021
-
ribosomal GTPase
-
-
-
-
ribosome-dependent GTPase
P39730
-
ribosome-dependent GTPase
Saccharomyces cerevisiae NOY891
P39730
-
-
ribosome-dependent GTPase
-
-
selenocysteine tRNA-specific elongation factor
-
-
signal recognition particle GTPase Ffh
-
-
SRP GTPase Ffh
-
-
SsEF-1alpha
P35021
amino acid at position 15, strain MT3 valine, SsMT3EF-1alpha, strain MT4 isoleucine, SsMT4EF-1alpha
SsGBP
Q980M3
-
-
SSO0269
Q980M3
locus name
SSO0269
Q980M3
locus name; locus name; locus name
-
SSO0412
Q97W59 and Q980A5 and Q97Z79
locus name, gamma-subunit
SSO0412
Q97W59 and Q980A5 and Q97Z79
locus name, gamma-subunit
-
SSO1050
Q97W59 and Q980A5 and Q97Z79
locus name, alpha-subunit
SSO1050
Q97W59 and Q980A5 and Q97Z79
locus name, alpha-subunit
-
SSO2381
Q97W59 and Q980A5 and Q97Z79
locus name, beta-subunit
SSO2381
Q97W59 and Q980A5 and Q97Z79
locus name, beta-subunit
-
SsoHflX
Q980M3
-
SsoHflX
Q980M3
-
-
translation factor aIF2/5B
-
-
translation initiation factor 2
-
-
translation initiation factor 2
Saccharomyces cerevisiae J293
-
-
-
translation initiation factor 2
Q980A5
-
translation initiation factor 2 gamma
-
-
translation initiation factor 2 gamma
Saccharomyces cerevisiae J293
-
-
-
translation initiation factor eIF5
-
-
translation initiation factor IF1
-
-
translation initiation factor IF2
-
-
translation termination factor eRF3
-
-
translational initiation factor 2
-
-
CAS REGISTRY NUMBER
COMMENTARY
9059-32-9
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Amphidinium carterae
-
-
-
Manually annotated by BRENDA team
-
P13905
UniProt
Manually annotated by BRENDA team
ecotype Landsberg erecta. Snowy cotyledon 1 mutant contains a mutation in a gene encoding the chloroplast elongation factor G, leading to an amino acid exchange within the predicted 70S ribosome-binding domain. The mutation results in a delay in the onset of germination. At this early developmental stage embryos still contain undifferntiated proplastids, whose proper function seems necessary for seed germination. In light-gropwn sco1 seedlings the greening of cotyledons is severely impaired, whereas the following true leaves develop normally as in wild-type plants
-
-
Manually annotated by BRENDA team
; strain DH5alpha
-
-
Manually annotated by BRENDA team
; used in hybrid system with Thermus thermophilus ribosomal protein L11 - EC 3.6.5.3
-
-
Manually annotated by BRENDA team
elongation factor G; elongation factor Tu
-
-
Manually annotated by BRENDA team
elongations factor (EF)Tu
-
-
Manually annotated by BRENDA team
elongations factor (EF)Tu; strain HW110, EF-Tu (wt) and EF-Tu (138N)
-
-
Manually annotated by BRENDA team
elongations factor (EF)Tu; strain MRE600, EF-Tu and EF-G
-
-
Manually annotated by BRENDA team
elongations factor (EF)Tu; strains MRE600, unmethylated enzyme, and LBE 1001, methylated enzyme
-
-
Manually annotated by BRENDA team
gene infB
-
-
Manually annotated by BRENDA team
strain B, elongation facor G ((EF)G)
-
-
Manually annotated by BRENDA team
strain MRE600, EF-Tu and EF-G
-
-
Manually annotated by BRENDA team
strain N4830, elongation factor EF-Tu (wt) and EF-Tu (D80N)
-
-
Manually annotated by BRENDA team
translation initiation factor IF2 mutants V400G and H448E, strain SL679R
-
-
Manually annotated by BRENDA team
Escherichia coli DH5alpha
strain DH5alpha
-
-
Manually annotated by BRENDA team
Escherichia coli HW110
strain HW110, EF-Tu (wt) and EF-Tu (138N)
-
-
Manually annotated by BRENDA team
Escherichia coli MRE600
-
-
-
Manually annotated by BRENDA team
Escherichia coli MRE600
strain MRE600, EF-Tu and EF-G
-
-
Manually annotated by BRENDA team
Escherichia coli N4830
strain N4830, elongation factor EF-Tu (wt) and EF-Tu (D80N)
-
-
Manually annotated by BRENDA team
elongation factor EF-1
-
-
Manually annotated by BRENDA team
expression in Escherichia coli
-
-
Manually annotated by BRENDA team
elongation factor (EF)Tu
-
-
Manually annotated by BRENDA team
Pleodorina sp.
-
-
-
Manually annotated by BRENDA team
strain 8830, elongation factor EF-Tu
-
-
Manually annotated by BRENDA team
Pseudomonas aeruginosa 8830
strain 8830, elongation factor EF-Tu
-
-
Manually annotated by BRENDA team
elongation factor (EF)Tu
-
-
Manually annotated by BRENDA team
eukaryotic elongation factor EF-2
-
-
Manually annotated by BRENDA team
eukaryotic initiaton factor eIF-2
-
-
Manually annotated by BRENDA team
Saccharomyces carlsbergensis EF-3
EF-3
-
-
Manually annotated by BRENDA team
strain NOY891
UniProt
Manually annotated by BRENDA team
Saccharomyces cerevisiae J293
strain J293
-
-
Manually annotated by BRENDA team
Saccharomyces cerevisiae NOY891
strain NOY891
UniProt
Manually annotated by BRENDA team
Schizosaccharomyces pombe EF-3
EF-3
-
-
Manually annotated by BRENDA team
elongation factor (EF)Tu
-
-
Manually annotated by BRENDA team
elongation factor (EF)Tu
-
-
Manually annotated by BRENDA team
elongation factor (EF)Tu
-
-
Manually annotated by BRENDA team
elongation factor (EF)Tu
-
-
Manually annotated by BRENDA team
; elongation factor 1alpha
-
-
Manually annotated by BRENDA team
; elongatipon factor 1alpha
-
-
Manually annotated by BRENDA team
elongation factor 1alpha from strain MT3 with optimum growth temperature 75C; elongation factor 1alpha from strain MT4 with optimum growth temperature 87C
-
-
Manually annotated by BRENDA team
elongation factor 2 exhibits an intrinsic hardly detectable GTPase activity that is stimulated by ribosomes up to 2000-fold
-
-
Manually annotated by BRENDA team
elongation factor SsEF-1alpha
-
-
Manually annotated by BRENDA team
Q97Z79: translation initiation factor 2 subunit alpha, Q97W59: translation initiation factor 2 subunit beta, Q980A5: translation initiation factor 2 subunit gamma
Q97W59 and Q980A5 and Q97Z79
UniProt
Manually annotated by BRENDA team
strain ATCC 49255, elongation factor aEF-1alpha
-
-
Manually annotated by BRENDA team
strain MT3, optimal growth temperature 75C, strain MT4, optimal growth temperature 87C
UniProt
Manually annotated by BRENDA team
Sulfolobus solfataricus ATCC 49255
-
-
-
Manually annotated by BRENDA team
Sulfolobus solfataricus MT-4
-
-
-
Manually annotated by BRENDA team
Q97Z79: translation initiation factor 2 subunit alpha, Q97W59: translation initiation factor 2 subunit beta, Q980A5: translation initiation factor 2 subunit gamma
Q97W59 and Q980A5 and Q97Z79
UniProt
Manually annotated by BRENDA team
elongation factor (EF)Tu
-
-
Manually annotated by BRENDA team
elongation factor (EF)Tu
-
-
Manually annotated by BRENDA team
strain JC469, JC496, JC 499, JC503
-
-
Manually annotated by BRENDA team
used in hybrid system with Escherichia coli ribosomal proteins L10, L11, L12 - EC 3.6.5.3
-
-
Manually annotated by BRENDA team
Thermus thermophilus JC469
strain JC469, JC496, JC 499, JC503
-
-
Manually annotated by BRENDA team
wheat germ translation factor 2 (WgeIF-2)
-
-
Manually annotated by BRENDA team
expression in Escherichia coli
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
an EF-G mutant lacking domains 4 and 5 shows ribosome-stimulated GTP hydrolysis activity 2.5fold slower than that of wild-type full-length EF-G and is insensitive to the effects of thiostrepton on both GTPase activity and ribosome binding
malfunction
Escherichia coli MRE600
-
an EF-G mutant lacking domains 4 and 5 shows ribosome-stimulated GTP hydrolysis activity 2.5fold slower than that of wild-type full-length EF-G and is insensitive to the effects of thiostrepton on both GTPase activity and ribosome binding
-
physiological function
-
the universally conserved GTPase HflX is a putative translation factor whose GTPase activity is stimulated by the 70S ribosome as well as the 50S but not the 30S ribosomal subunit
physiological function
-
elongation factor G, EF-G, is one of several GTP hydrolytic proteins that cycles repeatedly on and off the ribosome during protein synthesis in bacterial cells. In the functional cycle of EF-G, hydrolysis of GTP is coupled to tRNA-mRNA translocation in ribosomes. GTP hydrolysis induces conformational rearrangements in two switch elements in the G domain of EF-G and other GTPases. These switch elements are thought to initiate the cascade of events that lead to translocation and EF-G cycling between ribosomes, coupling mechanism, overview
physiological function
-
archaeal initiation factor 2 is a protein involved in the initiation of protein biosynthesis. In its GTP-bound, ON conformation, aIF2 binds an initiator tRNA and carries it to the ribosome. In its GDP-bound, OFF conformation, it dissociates from tRNA, molecular dynamics, overview. AIF2 is largely responsible for recruiting the first, initiator tRNA to the ribosome and positioning it correctly, in register with the start codon of the ribosome-bound mRNA
physiological function
-
the EF-G GTPase mediates the movement of the tRNA2-mRNA complex during translation
physiological function
-
EF-G and EF4 perform ribosome-dependent GTP hydrolysis and bind to conserved bases in 23S rRNA and stabilize ribosomal ratcheting
physiological function
-
importance of interdomain communication in IF2, importance of GTP as an IF2 ligand in the early initiation steps and the dispensability of the free energy generated by the IF2 GTPase in the late events of the translation initiation pathway
physiological function
-
protein synthesis requires several GTPase factors, including elongation factor Tu, EF-Tu, which delivers aminoacyl-tRNAs to the ribosome
physiological function
Escherichia coli MRE600
-
EF-G and EF4 perform ribosome-dependent GTP hydrolysis and bind to conserved bases in 23S rRNA and stabilize ribosomal ratcheting
-
metabolism
-
universal mechanism for GTPase activation and hydrolysis in translational GTPases on the ribosome
additional information
-
HflX-GTP exists in a structurally distinct 50S- and 70S-bound form that stabilizes GTP binding up to 70000fold and that may represent the GTPase-activated state. This activation is likely required for efficient GTP-hydrolysis, and may be similar to that observed in elongation factor G
additional information
-
cyclical movements of switch element I, sw1, within EF-G, Sw1 exposure depends on EF-G functional state, conformational changes in sw1 help to drive the unidirectional EF-G cycle during protein synthesis, intramolecular movements in EF-G, overview
additional information
-
protein:ligand interactions and conformational changes by molecular dynamics and Monte Carlo simulations, overview
additional information
-
IF2 mutant E571K, modified in its 30S binding domain IF2-G3, can perform in vitro all individual translation initiation functions of wild-type IF2 and supports faithful messenger RNA translation, despite having a reduced affinity for the 30S subunit and being completely inactive in GTP hydrolysis
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2',3'-O-N'-methylanthranilate-GTP + H2O
2',3'-O-N'-methylanthranilate-GDP + phosphate
show the reaction diagram
-
2',3'-O-N'-methylanthranilate, i.e. mant, is attached to GTP. EF-G binds and efficiently hydrolyzes mant-GTP in a ribosome-dependent manner
-
-
?
8-azido-GTP + H2O
8-azido-GDP + phosphate
show the reaction diagram
-
-
-
?
ATP + H2O
ADP + phosphate
show the reaction diagram
Saccharomyces carlsbergensis, Schizosaccharomyces pombe, Schizosaccharomyces pombe EF-3, Saccharomyces carlsbergensis EF-3
-
-
-
?
aurodox + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
Escherichia coli N4830
-
-
-
?
GDP + H2O
?
show the reaction diagram
Pseudomonas aeruginosa 8830
-
-
-
?
GDP + H2O
?
show the reaction diagram
Escherichia coli HW110
-
-
-
?
GTP + H2O
?
show the reaction diagram
-
70S ribosome, ribosome recycling factor, EF-G, GTP, 30C, 15 min
-
-
r
GTP + H2O
?
show the reaction diagram
P0CE48
kirromycin + H20
-
-
?
GTP + H2O
?
show the reaction diagram
Escherichia coli DH5alpha
-
70S ribosome, ribosome recycling factor, EF-G, GTP, 30C, 15 min
-
-
r
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
P32501
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
-
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
ir
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
P35021, -
-
-
-
ir
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
P39730
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
P60338
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Q01698
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
IF2 in complex with GTP, but not GDP promotes fast association of ribosomal subunits during initiation. IF2 promotes fast formation of the first peptide bond in the presence of GTP, but not GDP. GTP form of IF2 accelerates formation of the 70S ribosome from subunits and GTP hydrolysis accelerates release of IF2 from the 70S ribosome
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
importance of GTP hydrolysis in translation initiation for optimal cell growth
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
release of peptide promoted by the GGQ motif of class 1 release factors regulates the GTPase activity of RF3. Binding of GTP to RF3 and GTP hydrolysis requires peptide chain release
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
elongation factor G
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
elongation factor Tu
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
mutant of elongation factor G containing the effector loop from Thermus aquaticus EF-Tu has markedly decreased GTPase activity and does not catalyze translocation. The loops are not functionally interchangeable since the factors interact with different states of the ribosome
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
the catalytic role of His84 in elongation factor Tu is to stabilize the transition state of GTP hydrolysis by hydrogen bonding to the attacking water molecule or, possibly, the gamma-phosphate group of GTP
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
37C
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
elongation factor G catalyzes the translocation step in protein synthesis on the ribosome, enzyme-GTP and enzyme-GDP conformations in solution are very similar. The major contribution to the active GTPase conformation, which is quite different, therefore comes from its interaction with the ribosome
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
extodomain 2+3 stimulate the GTPase activity of ectodomain 1, extodomain 2+3 suppress the GTPase activity of ectodomain 1
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
GTPase activation due to C domain of the translation termination factor eRF1, which is bound with translation termination factor eRF3. As for the M and N domains, stimulation of eRF3 GTPase activity is more likely associated with the former, which is located in the large subunit along with the GTPase center of the ribosome, than with the latter, which is oriented towards the decoding center located in the small ribosomal subunit
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
the enzyme has the same domain structure and biochemical properties of a typical IF2 species as found in bacteria or mammalian mitochondria, but with enhanced ability to bind unformylated initiator met-tRNA
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
the integrity of the path between the peptidyltransferase center and both GTPase-associated center and sarcin-ricin loop is important for EF-G binding
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
the selenocysteine tRNA-specific elongation factor is responsible for the cotranslational incorporation of selenocysteine into proteins by recoding of a UGA step codon in the presence of a downstream mRNA hairpin loop
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
0.5 mM GTP, 37C, 10 min
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
60C
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
Base A 2660 is crucial for GTPase activity of EF-G. Reaction rates using reconstituted ribosomes, single turnover measurement, overview
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
EF-Tu is in its active conformation, when the switch I loop is ordered, and the catalytic histidine is coordinating the nucleophilic water in position for inline attack on the gamma-phosphate of GTP. The activated conformation is achieved due to a critical and conserved interaction of the histidine with A2662 of the sarcin-ricin loop of the 23S ribosomal RNA. Universal mechanism for GTPase activation and hydrolysis in translational GTPases on the ribosome. Premature GTP hydrolysis in EF-Tu is prevented by a hydrophobic gate consisting of residues Val20 of the P loop and Ile60 of switch I, which restricts access of His84 to the catalytic water
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
reaction using Escherichia coli 70S ribosomes, determination of binding of GTPases to 70S ribosomes in the GTP state, formation of 70S ribosome-tRNAPhe -GTPase-GDPNP complexes, multiple-turnover GTP hydrolysis
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
ribosome-dependent GTPase strongly stimulates the binding of initiator tRNA to the ribosomes even in the absence of other factors, aIF2/5B enhances the translation of both leadered and leaderless mRNAs when expressed in a cell-free protein-synthesizing system
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Q980M3
ATP hydrolysis is insignificant compared to the levels of GTP hydrolysis
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
displays either the intrinsic or the ribosome-dependent GTPase activity
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
slow GTPase with relatively low affinity for GTP
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Escherichia coli N4830
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Pseudomonas aeruginosa 8830
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Saccharomyces cerevisiae NOY891
P39730
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Schizosaccharomyces pombe EF-3, Saccharomyces carlsbergensis EF-3
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Escherichia coli HW110
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Sulfolobus solfataricus ATCC 49255
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Escherichia coli MRE600
-
-, reaction using Escherichia coli 70S ribosomes, determination of binding of GTPases to 70S ribosomes in the GTP state, formation of 70S ribosome-tRNAPhe -GTPase-GDPNP complexes, multiple-turnover GTP hydrolysis
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Escherichia coli MRE600
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Escherichia coli MRE600
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Q980M3
ATP hydrolysis is insignificant compared to the levels of GTP hydrolysis
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
slow GTPase with relatively low affinity for GTP
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Thermus thermophilus JC469
-
-
-
-
?
GTP gamma-(p-azido)anilide + H2O
GDP + phosphoric acid p-azidoanilin
show the reaction diagram
-
-
-
?
guanosine 5'-(thio)triphosphate + H2O
GDP + thiophosphate + 3 H+
show the reaction diagram
-
-
-
?
guanosine 5'-(thio)triphosphate + H2O
GDP + thiophosphate + 3 H+
show the reaction diagram
-
-
-
?
guanylyl imidodiphosphate + H2O
?
show the reaction diagram
-
-
-
?
guanylyl imidodiphosphate + H2O
?
show the reaction diagram
-
-
-
?
guanylyl imidodiphosphate + H2O
?
show the reaction diagram
-
-
-
?
ITP + H2O
IDP + phosphate
show the reaction diagram
-
-
-
?
XDP + H2O
XMP + phosphate
show the reaction diagram
Escherichia coli, Escherichia coli HW110
-
-
-
?
XTP + H2O
XDP + phosphate
show the reaction diagram
Escherichia coli, Escherichia coli HW110
-
-
-
?
ITP + H2O
IDP + phosphate
show the reaction diagram
Escherichia coli HW110
-
-
-
?
additional information
?
-
-
the enzyme exhibits significant binding activity with the nonformylated Met-tRNAf(Met)
-
-
-
additional information
?
-
-
eIF2A functions as a suppressor of Ure2p internal ribosome entry site-mediated translation in yeast cells
-
-
-
additional information
?
-
-
feeding artificial milk diets stimulate protein synthesis in skeletal muscle and liver of neonatal pigs by modulating the translation initiation factors that regulate mRNA binding to the ribosomal complex. However, provision of a high-protein diet that exceeds the protein requirement does not further enhance protein synthesis or translation initiator factor activation
-
-
-
additional information
?
-
Q980A5
aIF2 shows very high conformational flexibility in the alpha- and beta-subunits probably required for interaction of aIF2 with the small ribosomal subunit, overview
-
-
-
additional information
?
-
-
EF-1alpha shows GTPase activity and GDP-binding ability
-
-
-
additional information
?
-
-
Met-tRNA + 40S ribosomal subunit {?}
-
-
?
additional information
?
-
-
puromycin + 50S subunit {?}
-
-
?
additional information
?
-
-
EF4-ribosome interactions during reverse translocation, overview
-
-
-
additional information
?
-
-
HflX interacts with 50S and 70S particles, and also with the 30S subunit, independent of the nucleotide-bound state and in tight binding, minimal model for the functional cycle of HflX, interaction with the 70S ribosome and functional mechanism of HflX, overview
-
-
-
additional information
?
-
-
residue 196 is located in a solvent-exposed location of the G' subdomain, while its neighboring helices AG' and BG' make contacts with protein L7/L12 of the ribosome. The latter contacts involve conserved electrostatically interacting residues that allosterically activate GTP hydrolysis in the G domain of EF-G. Residue 58 moves substantially from its initial ordered position adjacent to helix BIII
-
-
-
additional information
?
-
-
structure-activity relationship, molecular dynamics simulations, overview
-
-
-
additional information
?
-
Saccharomyces cerevisiae J293
-
Met-tRNA + 40S ribosomal subunit {?}
-
-
?
additional information
?
-
Escherichia coli MRE600
-
EF4-ribosome interactions during reverse translocation, overview
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + H2O
ADP + phosphate
show the reaction diagram
Saccharomyces carlsbergensis, Schizosaccharomyces pombe, Schizosaccharomyces pombe EF-3, Saccharomyces carlsbergensis EF-3
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
-
-
-
?
GDP + H2O
?
show the reaction diagram
Escherichia coli N4830
-
-
-
?
GDP + H2O
?
show the reaction diagram
Pseudomonas aeruginosa 8830
-
-
-
?
GDP + H2O
?
show the reaction diagram
Escherichia coli HW110
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
ir
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
P35021, -
-
-
-
ir
GTP + H2O
GDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
IF2 in complex with GTP, but not GDP promotes fast association of ribosomal subunits during initiation. IF2 promotes fast formation of the first peptide bond in the presence of GTP, but not GDP. GTP form of IF2 accelerates formation of the 70S ribosome from subunits and GTP hydrolysis accelerates release of IF2 from the 70S ribosome
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
importance of GTP hydrolysis in translation initiation for optimal cell growth
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
release of peptide promoted by the GGQ motif of class 1 release factors regulates the GTPase activity of RF3. Binding of GTP to RF3 and GTP hydrolysis requires peptide chain release
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
elongation factor G catalyzes the translocation step in protein synthesis on the ribosome
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
-
ribosome-dependent GTPase strongly stimulates the binding of initiator tRNA to the ribosomes even in the absence of other factors
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Escherichia coli N4830
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Pseudomonas aeruginosa 8830
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Schizosaccharomyces pombe EF-3, Saccharomyces carlsbergensis EF-3
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Escherichia coli HW110
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Escherichia coli MRE600
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Escherichia coli MRE600
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
Escherichia coli MRE600
-
-
-
?
guanosine 5'-(thio)triphosphate + H2O
GDP + thiophosphate + 3 H+
show the reaction diagram
-
-
-
?
guanosine 5'-(thio)triphosphate + H2O
GDP + thiophosphate + 3 H+
show the reaction diagram
-
-
-
?
guanylyl imidodiphosphate + H2O
?
show the reaction diagram
-
-
-
?
guanylyl imidodiphosphate + H2O
?
show the reaction diagram
-
-
-
?
guanylyl imidodiphosphate + H2O
?
show the reaction diagram
-
-
-
?
ITP + H2O
IDP + phosphate
show the reaction diagram
-
-
-
?
XDP + H2O
XMP + phosphate
show the reaction diagram
Escherichia coli, Escherichia coli HW110
-
-
-
?
XTP + H2O
XDP + phosphate
show the reaction diagram
Escherichia coli, Escherichia coli HW110
-
-
-
?
ITP + H2O
IDP + phosphate
show the reaction diagram
Escherichia coli HW110
-
-
-
?
additional information
?
-
-
eIF2A functions as a suppressor of Ure2p internal ribosome entry site-mediated translation in yeast cells
-
-
-
additional information
?
-
Escherichia coli, Escherichia coli MRE600
-
EF4-ribosome interactions during reverse translocation, overview
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
BaCl2
-
10 mM, about 8fold stimulation
CaCl2
-
10 mM, highest stimulation by BaCl2 (8fold), followed by SrCl2, MgCl2, MnCl2, CaCl2 and CoCl2 in a decreasing order of effectiveness
CoCl2
-
10 mM, highest stimulation by BaCl2 (8fold), followed by SrCl2, MgCl2, MnCl2, CaCl2 and CoCl2 in a decreasing order of effectiveness
Mg2+
-
GTPase bound to Mg2+GDP reveals two new binding conformations. In the first the protein undergoes a conformational change that brings a conserved aspartate into its second coordination sphere. In the second, the magnesium coordination sphere is disrupted so that only five oxygen ligands are present
Mg2+
-
plays a marginal role in the nucleotide exchange process
Mg2+
-
required
Mg2+
-
required
Mg2+
-
requires Mg2+ for its full GTPase catalytic activity
MgCl2
-
10 mM, highest stimulation by BaCl2 (8fold), followed by SrCl2, MgCl2, MnCl2, CaCl2 and CoCl2 in a decreasing order of effectiveness
NaCl
-
GTPase activity is measured in the presence of 3.6 M NaCl
SrCl2
-
10 mM, highest stimulation by BaCl2 (8fold), followed by SrCl2, MgCl2, MnCl2, CaCl2 and CoCl2 in a decreasing order of effectiveness
Zn2+
-
zinc-binding domain in the beta-subunit
MnCl2
-
10 mM, highest stimulation by BaCl2 (8fold), followed by SrCl2, MgCl2, MnCl2, CaCl2 and CoCl2 in a decreasing order of effectiveness
additional information
P13905
decreased gene expression of A1, A2, A3 and A4 gene under iron deficiency
additional information
-
GTPase stimulated by ethylene glycol and BaCl2 does not require the presence of univalent cations. Li+, Na+, K+ or NH4+ added singularly up to 1 M concentration, do not produce any significant stimulation of SsEF-2 GTPase either in the absence or in the presence of ethylene glycol. They reduce the stimulation of SsEF-2 GTPase by ethylene glycol plus BaCl2 or SrCl2
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Anti-EF-Tu antibody
-
-
-
Anti-Ndk antibody
-
-
-
Chloramphenicol
-
-
dihydrostreptomycin
-
-
EF-G GTPase inhibitor
-
-
-
Fusidic acid
-
inhibition of ribosome disassembly by EF-G and ribosome recycling factor, no influence on GTP hydrolysis
GDP
-
competitive with GTP
GDP
-
GTPase activity in the presence of a molar concentration of NaCl is competitively inhibited
GDP gamma-S
-
-
GE2270A
-
antibiotic inhibits intrinsic GTPase and that stimulated by ribosomes; thiazolyl-peptide antibiotic, inhibits both the intrinsic GTPase of elongation factor 1alpha and that stimulated by ribosomes. The M domain is the region of the enzyme most responsible for the interaction with GE2270A
guanidine hydrochloride
-
deactivation by denaturation of the protein
guanosine 5'-(beta,gamma-imido)triphosphate
-
-
guanosine 5'-tetraphosphate
-
competitive inhibition of intrinsic GTPase, inhibition of archaeal protein synthesis in vitro, even though the concentration required to get inhibition is higher than that required for the eubacterial and eukaryal systems
guanosine-5'-[(beta,gamma)-imido]triphosphate
-
i.e. GppNHp. GTPase activity in the presence of a molar concentration of NaCl is competitively inhibited
-
guanyl-5'-yl imidodiphosphate
-
competitive with GTP
guanyl-5'-yl imidotriphosphate
-
-
guanyl-5'yl-imidodiphosphate
-
-
hygromycin A
-
-
N-ethylmaleimide
-
-
NH4Cl
-
at higher concentration
P3-1-(2-nitro)phenylethylguanosine 5'-O-triphosphate
-
-
pulvomycin
-
the antibiotic is able to reduce in vitro the rate of protein synthesis however, the concentration of pulvomycin leading to 50% inhibition (173 mM) is two order of magnitude higher but one order lower than that required in eubacteria and eukarya, respectively. Pulvomycin is able to decrease the affinity of the elongation factor toward aa-tRNA only in the presence of GTP, to an extent similar to that measured in the presence of GDP
ribostamycin
-
-
sparsomycin
-
-
spermidine
-
2mM
streptogramin A
-
-
tetracyclin
-
-
tetracycline
-
mixed inhibition. The inhibition level depends on the antibiotic concentration, even though a complete inhibition is not reached even in the presence of 0.120 mM antibiotic, a concentration corresponding to about 200fold molar excess over the elongation factor
Thiostrepton
-
inhibits the ribosome-stimulated GTPase activity of EF-G and EF4. An EF-G mutant lacking domains 4 and 5 is insensitive to the effects of thiostrepton on both GTPase activity and ribosome binding
translation initiation factor IF1
-
-
-
translation initiation factor IF3
-
inhibition could be overcome by increasing concentrations of divalent cations
-
Urea
-
deactivation by denaturation of the protein
vanadate
-
inhibition of ribosome disassembly, no influence on GTP hydrolysis
additional information
P32501
subunit alpha, beta and delta of eIF2B down-regulates activity of the eIF2B catalytic subcomplex
-
additional information
-
viomycin and fusidic acid do not prevent GTP hydrolysis, but these antibiotics trap EF-G on the ribosome before or after ribosomal translocation, respectively
-
additional information
-
ribosomes lacking the 23S rRNA and with deletion of SRL region, but not of GAC, inactivate EF-G1
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1-Propanol
-
stimulation by aliphatic alcohols in a decreasing order of effectiveness: ethylene glycol > 2-propanol > ethanol > glycerol > methanol > 1-propanol
2-Propanol
-
stimulation by aliphatic alcohols in a decreasing order of effectiveness: ethylene glycol > 2-propanol > ethanol > glycerol > methanol > 1-propanol
bluensomycin
-
-
ethanol
-
stimulation by aliphatic alcohols in a decreasing order of effectiveness: ethylene glycol > 2-propanol > ethanol > glycerol > methanol > 1-propanol
ethylene glycoI
-
60%, 300fold stimulation
-
gentamicin C1
-
-
gentamicin C1a
-
-
glycerol
-
stimulation by aliphatic alcohols in a decreasing order of effectiveness: ethylene glycol > 2-propanol > ethanol > glycerol > methanol > 1-propanol
kanamycin A
-
-
kanamycin B
-
-
kirromycin
-
enhances activity of mutant enzyme G13A (maximal stimulation at 0.04 mM), does not stimulate intrinsic GTPase of SsEF-1alpha triggered by 3.6 M NaCl
L7/12
-
functional compatibility between elongation factor G and the L7/12 protein in the ribosome governs its translational specificity; the C-terminal domian of L7/12 is responsible for EF-Tu function. Functional compatibility between elongation factor Tu and the L7/12 protein in the ribosome governs its translational specificity
-
NaCl
-
intrinsic GTPase activity of elongation factor 1alpha is triggered in vitro by NaCl at molar concentrations. The sodium ion is responsible for the induction of the GTPase activity, whereas the anion modulates the enzymatic activity through destabilization of particular regions of the protein; intrinsic GTPase activity that is triggered in vitro by NaCl at molar concentrations
pulvomycin
-
increasing pulvomycin concentration increased the rate of the intrinsic GTPase catalysed by elongation factor 1alpha, reaching its maximum stimulation effect at 30 mM. Pulvomycin exerts its stimulatory function at all the tested temperatures (45-75C).
ribosomal subunits
-
-
-
ribosomal subunits
-
-
-
ribosome
-
stimulates GTPase activity of elongation factor Tu. The factor binding site is loacetd on the 50S ribosomal subunit and comprises proteins L7/12, L10, L11, the l11-binding region of 23 rRNA, and the sarcin-ricin loop of 23S rRNA. L7/12 stimulates the GTPase activity of elongation factor G by inducing the catalytically active conformation of the G domain; stimulates GTPase activity of elongation factor Tu. The factor binding site is loacetd on the 50S ribosomal subunit and comprises proteins L7/12, L10, L11, the l11-binding region of 23 rRNA, and the sarcin-ricin loop of 23S rRNA. L7/12 stimulates the GTPase activity of elongation factor Tu by inducing the catalytically active conformation of the G domain
-
streptomycin
-
-
streptomycin
-
only EF-G
methanol
-
stimulation by aliphatic alcohols in a decreasing order of effectiveness: ethylene glycol > 2-propanol > ethanol > glycerol > methanol > 1-propanol
additional information
P32501
active in GTP-bound form, inactive in GDP-bound form; phosphorylation
-
additional information
Q01698
active in GTP-bound form, inactive in GDP-bound form
-
additional information
P60338
active in GTP-bound form, inactive in GDP-bound form
-
additional information
-
GTP hydrolysis is essential for release of eIF5B from the 80S ribosomal subunit
-
additional information
P0CE48
activity triggered by ribosome-induced conformational changes of EF-Tu
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0008
-
GTP
-
wild type
0.0008
-
GTP
-
mutant A26G, pH 7.5, 60C, presence of 40% ethylene glycol
0.0009
-
GTP
P35021, -
SsMT4EF-1alpha
0.0009
-
GTP
-
pH 7.8, 50C
0.001
-
GTP
-
wild-type, pH 7.5, 60C, presence of 40% ethylene glycol
0.001
-
GTP
-
pH 7.8, 75C
0.0012
-
GTP
-
pH 7.8, 60C
0.0013
-
GTP
-
pH 7.8, 80C
0.0014
-
GTP
-
pH 7.8, 70C
0.0018
-
GTP
-
mutant D60A
0.002
0.009
GTP
-
depending on NaCl-concentration and temperature
0.0023
-
GTP
-
Ss(GM)EF-1alpha; truncated mutant, domains G+M, 60C, pH 7.8
0.0024
-
GTP
-
Ss(G)EF-1alpha; truncated mutant, domain G, 60C, pH 7.8
0.0025
-
GTP
-
SsEF-1alpha; wild-type, 60C, pH 7.8
0.0025
-
GTP
-
pH 7.8, 60C
0.0027
-
GTP
-
60C, wild-type enzyme
0.0027
-
GTP
P35021, -
SsMT3EF-1alpha
0.00436
-
GTP
-
pH 7.8, 50C, GTPase activity in the presence 3.6 M NaCl
0.0046
-
GTP
-
60C, mutant enzyme G13A
0.0053
-
GTP
-
pH 7.8, 87C
0.0053
-
GTP
-
pH 7.4, 50C, in absence of ribosomes
0.0053
-
GTP
-
pH 7.5, 50C, wild-type enzyme
0.0055
-
GTP
-
mutant A26G, pH 7.5, 60C, presence of 10% ethylene glycol
0.0075
-
GTP
-
pH 7.5, 60C
0.008
-
GTP
-
pH 7.8, 50C
0.009
-
GTP
-
pH 7.8, 60C
0.0096
-
GTP
-
wild-type, pH 7.5, 60C, presence of 10% ethylene glycol
0.01
-
GTP
-
pH 7.8, 70C
0.011
-
GTP
-
pH 7.8, 75C
0.011
-
GTP
-
pH 7.5, 50C, mutant enzyme F236P
0.0129
-
GTP
-
pH 7.8, 50C, N-terminal deletion mutant
0.013
-
GTP
-
pH 7.8, 80C
0.0141
-
GTP
-
pH 7.8, 50C, full-length enzyme
0.0153
-
GTP
-
V400G
0.016
-
GTP
-
D80N
0.0194
-
GTP
-
mutant A26G, pH 7.5, 60C
0.02
-
GTP
-
pH 7.8, 91C
0.0247
-
GTP
-
H448E
0.03
-
GTP
-
wild type
0.032
-
GTP
-
pH 7.8, 87C
0.0532
-
GTP
-
pH 7.8, 95C
0.148
-
GTP
-
pH 7.8, 91C
additional information
-
additional information
-
substrate and product binding kinetics and thermodynamics, overview
-
additional information
-
additional information
-
pre-steady-state kinetic analysis of HflX and Hflx ribosomal complexes interacting with GDP and a nonhydrolyzable analogue of mant-GTP, overview
-
additional information
-
additional information
-
ribosome binding kinetics of GTP hydrolysis-inactive recombinant EF-G mutants 58C and 196C labeled with 2',7'-difluorofluorescein maleimide, i.e. 58C-mant and 196C-mant, overview
-
additional information
-
additional information
-
substrate and product binding kinetics and thermodynamics with ON and OFF aIF2 , overview
-
additional information
-
additional information
-
substrate binding kinetics of wild-type and mutant IF2s, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0004
-
GTP
-
pH 7.5, 50C, wild-type enzyme
0.00053
-
GTP
-
pH 7.5, 50C, mutant enzyme F236P
0.00092
-
GTP
-
pH 7.4, 50C, in absence of ribosomes
0.001
-
GTP
-
Ss(G)EF-1alpha
0.00105
-
GTP
-
pH 7.8, 50C, full-length enzyme
0.00167
-
GTP
-
60C, mutant enzyme G13A
0.003
-
GTP
-
pH 7.8, 50C, GTPase activity in the presence 3.6 M NaCl
0.0033
-
GTP
-
wild-type, pH 7.5, 60C, presence of 10% ethylene glycol
0.01
-
GTP
-
truncated mutant, domain G, 60C, pH 7.8
0.013
-
GTP
-
60C, wild-type enzyme
0.013
-
GTP
P35021, -
SsMT3EF-1alpha
0.014
-
GTP
-
SsEF-1alpha; wild-type, 60C, pH 7.8
0.015
-
GTP
P35021, -
SsMT4EF-1alpha
0.018
-
GTP
-
Ss(GM)EF-1alpha; truncated mutant, domains G+M, 60C, pH 7.8
0.023
-
GTP
-
pH 7.8, 50C
0.026
-
GTP
-
pH 7.8, 50C, N-terminal deletion mutant
0.032
-
GTP
-
wild-type, pH 7.5, 60C, presence of 40% ethylene glycol
0.038
-
GTP
-
pH 7.8, 60C
0.118
-
GTP
-
pH 7.8, 70C
0.12
-
GTP
-
mutant A26G, pH 7.5, 60C, presence of 10% ethylene glycol
0.133
-
GTP
-
mutant A26G, pH 7.5, 60C
0.152
-
GTP
-
pH 7.8, 75C
0.165
-
GTP
-
mutant A26G, pH 7.5, 60C, presence of 40% ethylene glycol
0.2
-
GTP
-
pH 7.8, 50C
0.23
-
GTP
-
pH 7.8, 80C
0.5
-
GTP
-
pH 7.8, 60C
0.512
-
GTP
-
pH 7.8, 87C
0.558
-
GTP
-
pH 7.8, 95C
0.8
-
GTP
-
pH 7.8, 60C
0.89
-
GTP
-
pH 7.8, 91C
0.9
-
GTP
-
pH 7.5, 60C
1.7
-
GTP
-
pH 7.8, 70C
2.7
-
GTP
-
pH 7.8, 75C
3.5
-
GTP
-
pH 7.8, 80C
6.2
-
GTP
-
pH 7.8, 87C
8.8
-
GTP
-
pH 7.8, 91C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.048
-
GTP
-
pH 7.5, 50C, mutant enzyme F236P
11186
0.076
-
GTP
-
pH 7.5, 50C, wild-type enzyme
11186
0.17
-
GTP
-
pH 7.4, 50C, in absence of ribosomes
11186
0.69
-
GTP
-
pH 7.8, 50C, GTPase activity in the presence 3.6 M NaCl
11186
11
-
GTP
-
pH 7.8, 95C
11186
26
-
GTP
-
pH 7.8, 50C
11186
32
-
GTP
-
pH 7.8, 60C
11186
45
-
GTP
-
pH 7.8, 91C
11186
84
-
GTP
-
pH 7.8, 70C
11186
97
-
GTP
-
pH 7.8, 87C
11186
152
-
GTP
-
pH 7.8, 75C
11186
177
-
GTP
-
pH 7.8, 80C
11186
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00012
-
GDP
-
pH 7.8, 50C, GTPase activity in the presence 3.6 M NaCl
0.0003
-
GDP
-
wild-type, pH 7.5, 60C, presence of 40% ethylene glycol
0.0005
-
GDP
-
mutant A26G, pH 7.5, 60C, presence of 40% ethylene glycol
0.0008
-
GDP
-
60C, wild-type enzyme
0.0013
-
GDP
-
SsEF-1alpha; wild-type, 60C, pH 7.8
0.0017
-
GDP
-
Ss(G)EF-1alpha; truncated mutant, domain G, 60C, pH 7.8
0.0029
-
GDP
-
mutant D60A
0.0042
-
GDP
-
60C, mutant enzyme G13A
0.0062
-
GDP
-
Ss(GM)EF-1alpha; truncated mutant, domains G+M, 60C, pH 7.8
0.0074
-
GDP
-
mutant A26G, pH 7.5, 60C
0.0084
-
guanosine 5'-(beta,gamma-imido)triphosphate
-
SsEF-1alpha
0.0193
-
guanosine 5'-(beta,gamma-imido)triphosphate
-
Ss(G)EF-1alpha
0.0407
-
guanosine 5'-(beta,gamma-imido)triphosphate
-
Ss(GM)EF-1alpha
0.00047
-
guanosine-5'-[(beta,gamma)-imido]triphosphate
-
pH 7.8, 50C, GTPase activity in the presence 3.6 M NaCl
-
0.0084
-
guanyl-5'-yl imidotriphosphate
-
wild-type, 60C, pH 7.8
0.0193
-
guanyl-5'-yl imidotriphosphate
-
truncated mutant, domain G, 60C, pH 7.8
0.0407
-
guanyl-5'-yl imidotriphosphate
-
truncated mutant, domains G+M, 60C, pH 7.8
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0059
-
GE2270A
-
half-inhibition of the GTPase of the isolated domain GM of SsEF-1alpha
0.007
-
GE2270A
-
half-inhibition of the NaCl-dependent GTPase of SsEF-1alpha
0.011
-
GE2270A
-
half-inhibition of the ribosome-dependent GTPase of SsEF-1alpha
0.0148
-
GE2270A
-
half-inhibition of the GTPase of the isolated domain G of SsEF-1alpha
0.0463
-
GE2270A
-
half-inhibition of the GTPase of a chimeric EF containing the domain G of SsEF-1alpha and the domains MC of Escherichia coli EF-Tu
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.4
-
-
assay at
7.4
-
-
assay at
7.5
-
-
assay at
7.5
-
-
assay at
7.8
-
-
ATPase assay
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
-
-
assay at
32
-
-
GTPase activity of domain 1
37
-
-
assay at
37
-
-
assay at
38
-
-
GTPase activity of domain 1 fused to domain 2+3
50
-
-
GTPase activity of domain 1
60
-
-
activity assay
60
-
-
ATPase assay
60
-
-
the GTPase activity is measured in the presence of either 3.6 M NaCl or 1.6 microM ribosomes
61
-
-
GTPase activity of domain 1 fused to domain 2+3
80
-
-
mutant G13A
80
-
-
both Km for GTP and kcat of GTPaser increase with increasing temperature
87
-
P35021, -
SsMT3EF-1alpha
90
-
-
wild-type enzyme
91
-
P35021, -
SsMT4EF-1alpha
additional information
-
-
the rate of nucleotide binding to aEF-1 a increased with temperature, reaching a maximum at 95C
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
70
86
-
about 60% of maximal activity at 70C and at 86C, mutant enzyme G13A
70
95
-
70C: about 60% of maximal activity, 95C: about 90% of maximal activity, wild-type enzyme
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
9.1
-
-
isoelectric focusing, pH-gradient 3-10
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
EF-G1mt is active on both bacterial and mitochondrial ribosomes
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Halobacterium salinarum (strain ATCC 29341 / DSM 671 / R1)
Methanosarcina mazei (strain ATCC BAA-159 / DSM 3647 / Goe1 / Go1 / JCM 11833 / OCM 88)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
28000
-
-
truncated G-form of SsEF-1alpha, determined by SDS-PAGE and Western blot
38000
-
-
truncated GM-form of SsEF-1alpha, determined by SDS-PAGE and Western blot
43000
-
-
-
45000
-
-
SDS-PAGE
49000
-
-
gel filtration; SDS-PAGE
49000
-
-
gel filtration
50000
-
-
SDS-PAGE
58000
-
-
SDS-PAGE
95000
-
-
gel filtration
120000
-
-
SDS-PAGE
125000
-
-
SDS-PAGE
150000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 50500, His6-tagged Hflx, SDS-PAGE
?
-
x * 70000, His-tagged protein, SDS-PAGE
heteropentamer
P32501
alphabetagammadeltaepsilon
monomer
-
1*49000, SDS-PAGE and gel filtration
monomer
-
1 * 49000, SDS-PAGE
pentamer
-
-
tetramer
-
1 * 37000 + 1 * 40000 + 1 * 42000 + 1 * 52000, SDS-PAGE
tetramer
-
1 * 50000 + 1 * 30000 + 1 * 52000 + 1 * 40000
tetramer
Schizosaccharomyces pombe EF-3
-
1 * 37000 + 1 * 40000 + 1 * 42000 + 1 * 52000, SDS-PAGE
-
trimer
-
1 * 32000 + 1 * 35000 + 1 * 55000, sedimentation equilibrium centrifugation
trimer
-
heterotrimer, the archaeal factor aIF2 is formed upon the 1:1:1 association of three subunits: alpha, beta, and gamma
trimer
P32501
alphabetagamma
monomer
Sulfolobus solfataricus MT-4
-
1 * 49000, SDS-PAGE
-
additional information
-
aIF2 shows very high conformational flexibility in the alpha- and beta-subunits probably required for interaction of aIF2 with the small ribosomal subunit, structure comparison of the heterotrimer aIF2alphabetagamma to aIF2alphagamma and aIF2betagamma heterodimers, structure of subunits, overview
additional information
-
structure-activity relationship, molecular dynamics simulations, overview
additional information
-
the enzyme binds to the 50S ribosomal subunit
additional information
-
the enzyme binds to the 50S ribosomal subunit
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-
trypsin degradation
side-chain modification
-
methylation of lysine-56, enzyme with decreased rate of tRNA-dependent GTP hydrolysis
side-chain modification
-
acetylation and methylation
proteolytic modification
-
chymotrypsin degradation
phosphoprotein
-
recombinant subunits of eIF2alpha and beta-subunits are also phosphorylated in cultured insect cells. Phosphorylation of eIF2alpha in vitro is not significantly different in the presence and absence of the other subunits
phosphoprotein
-
phosphorylation of the alpha-subunit of the eukaryotic initiation factor-2 (eIF2alpha) reduces protein synthesis and enhances apoptosis in response to proteasome inhibition
phosphoprotein
-
heme-regulated inhibitor kinase-mediated phosphorylation of eukaryotic translation initiation factor 2 inhibits translation, induces stress granule formation, and mediates survival upon arsenite exposure
side-chain modification
-
phosphorylation of alpha, beta and gamma subunits of EF-1, phosphorylated EF-1 with increased GDP/GTP-exchange activity
proteolytic modification
-
chymotrypsin degradation
proteolytic modification
-
trypsin degradation, four fragmentation products: 1 * 82000 + 1 * 48000 + 1 * 33000-34000 + 1 * 10000, SDS-PAGE
proteolytic modification
-
trypsin degradation
proteolytic modification
-
trypsin degradation
side-chain modification
-
acetylation and methylation
proteolytic modification
-
trypsin degradation
side-chain modification
-
acetylation and methylation
side-chain modification
-
acetylation and methylation
additional information
-
differently from the wild-type enzyme the recombinant enzyme does not undergo post-translational modification of His603 into diphthamide, as indicated by its inability to be ADP-ribosylated
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
sitting drop vapor diffusion method, crystal structure of elongation factor Tu*Ts complex at 2.2 A resolution
-
two crystal forms of a complex between trypsin-modified elongation factor Tu-MgGDP and the antibiotic tetracycline solved by X-ray diffraction analysis to resolution of 2.8 and 2.1 A, respectively
-
hanging-drop vapour-diffusion method
-
vapour-diffusion method from ammonium sulfate either in the presence of GDP, GppHNp or without nucleotide, yielding isomorphous crystals for all three forms
-
crystal structure of the regulatory subunit aIF2Balpha, hanging-drop vapour diffusion method at 20C, three-dimensional structure is determined by X-ray crystallography at 2.2 A resolution
-
5 A resolution crystal structure of the ternary complex formed by archaeal aIF2 from Sulfolobus solfataricus, the GTP analog GDPNP and methionylated initiator tRNA
Q97W59 and Q980A5 and Q97Z79
analysis of crystal structures of ON and OFF aIF2 at resolution of 3.0 and 2.15 A
-
crystal structure analysis, overview
-
crystal structure of HflX from Sulfolobus solfataricus solved to 2.0 A resolution in apo- and GDP-bound forms
-
crystals of wild-type enzyme/GDP comple and mutant enzymes G235P and G235S are grown using hanging drop method at 16C
-
elongation factor 1alpha in complex with GDP, structure at 1.8 A resolution
-
elongation factor 1alpha in complex with Mg2+ (100 mM) and GDP. Elongation factor 1alpha in complex with GDP does not bind Mg2+, when the ion is present in the crystallization medium at moderate concentrations (5 mM). Crystals are grown using PEG 4000 and propan-2-ol as precipitants. Diffraction quality crystals are obtained using microbatch under oil technique at 4C and a protein concentration of 6 mg/ml
-
Fourier transform infrared spectroscopic study. Substitution of the GDP bound with guanyl-5'-yl imido diphosphate induces a slight increase in the alpha helix and beta sheet content. The alpha helix content of the enzyme-GDP complex increases upon addition of salts, and the highest effect is produced by 5 M NaCl. Thermal stability of the enzyme-GDP complex is significantly reduced when the GDP is replaced with guanyl-5'-yl imido diphosphate or in the presence of NaBr or NH4Cl
-
full-sized alphabetagamma heterotrimeric aIF2 in the nucleotide-free form, and aIF2alphagamma dimer, X-ray diffraction structure determination and analysis at 2.8 A resolution
-
crystal structure of the Mg2+-GDP complex of the Ffh NG-domain refined at 2.1 resolution
-
EF-Tu bound to aminoacyl-tRNA of the 70s ribosome and a GTP analogue, X-ray diffraction structure determination and analysis at 3.1 A resolution
-
structure of the mutant enzyme T84A in complex with the non-hydrolysable GTP analogue GDPNP
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
100
-
EF-1alpha denaturation profile at pH 4.0, the protonation state of the numerous Asp and Glu residues plays a critical role for the thermally denatured state of the enzyme, reversibility of the inter-conversion between the two denatured forms, overview
87
96
-
half-inactivation time 99 min at 87C, half-inactivation time 3.5 min at 96C
87
-
-
half-denaturation of mutant enzyme G13A
91
-
-
10 min, 50% inactivation of GDP binding ability, mutant enzyme G13A
92
-
-
half-denaturation of wild-type enzyme
93.6
-
-
temperature for half denaturation of D60A mutant
94
-
-
10 min, 50% inactivation of GDP binding ability, wild-type enzyme
95
-
-
10 min, 50% inactivation
95.1
-
-
temperature for half denaturation
96
-
-
binding of guanosine 5'-tetraphosphate to SsEF-1alpha renders the elongation factor more resistant to heat treatment. The denaturation profile of the elongation factor in the presence of guanosine tetraphosphate is shifted towards higher temperatures with a denaturation midpoint (96.4C) about 2C higher with respect to that observed for the elongation factor bound to GDP
additional information
-
-
domain 1 and domains 2+3 of both EF-Tu positively cooperate to heat-stabilize the GTPase center to attain optimal activity at alpha-helical regions of the G-domain
additional information
-
-
thermal stability of the enzyme-GDP complex is significantly reduced when the GDP is replaced with guanyl-5'-yl imido diphosphate or in the presence of NaBr or NH4Cl
additional information
-
-
after a 6 h treatment at 80C and 87C, the residual activity is 80% and 54%, respectively; at 95C the [3H]GDP-binding activity became one half of the initial activity after about 30 min. After 80 days at 25C, a crude aEF-la preparation retained 50% of the maximum nucleotide-binding capacity
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
EF-Tu(138N) not functional in vivo
-
mutants lethal to E. coli
-
eIF2A is an inherently unstable protein with a half-life of about 17 min
-
conformational change of the elongation factor takes place upon interaction with the antibiotic. Protection against chemical denaturation of SsEF-1alpha is observed in the presence of pulvomycin
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Glycerol
-
-
Glycerol
Escherichia coli N4830
-
-
-
guanidine-HCl
-
50% inactivation of GTPase activity at 3.0 M guanidine-HCl in presence 3.6 M NaCl and absence of pulvomycin. 50% inactivation of GTPase activity at 3.6 M guanidine-HCl in presence 3.6 M NaCl and presence of pulvomycin
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, 20 mM Tris/HCl, pH 7.8, 50 mM KCl, 10 mM MgCl2, 50% v/v glycerol
-
-20C, stable for several months
-
-20C, Tris/HCl buffer, pH 7.8, 10 mM MgCl2, 50 mM KCl, 50% v/v glycerol
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
high level expression of mutant enzyme P269S in Escherichia coli
-
expression in Escherichia coli BL21
-
mutant enzyme Q290L, expression in Escherichia coli
-
recombinant expression of EF-G mutants 58C and 196C
-
expression in Escherichia coli
-
overexpression of all three subunits of human eIEF2 independently, and together in Sf9 cells using pFast Bac HT vector of baculovirus expression system. the expression of all subunits increases in infection time up to 72 h. Expression of the mutant forms S51A, S51D and S48A
-
full-length enzyme or C2 subdomain of ymIF2 expressed in Escherichia coli. The full-length ymIF2 can substitute for Escherichia coli IF2 in the formation of a functional initiation complex on 70S Escherichia coli ribosomes capable of forming the first peptide bond
-
His-tagged version of ymIF2 lacking its predicted mitochondrial presequence is expressed in Escherichia coli
-
comparison of sequence with that of Sulfolobus solfataricus strain MT4 shows only one amino acid change, i.e. I15V. The difference is in the first guanine nucleotide binding consensus sequence G13HIDHGK and is responsible for a increased efficiency in protein synthesis, which is accompanied by an reduced affinity for both guanosine diphosphate (GDP) and guanosine triphosphate (GTP), and an decreased efficiency in the intrinsic GTPase activity; comparison of sequence with that of Sulfolobus solfataricus strain MT4 shows only one amino acid change, i.e. V15I. The difference is in the first guanine nucleotide binding consensus sequence G13HIDHGK and is responsible for a reduced efficiency in protein synthesis, which is accompanied by an increased affinity for both guanosine diphosphate (GDP) and guanosine triphosphate (GTP), and an increased efficiency in the intrinsic GTPase activity; the gene encoding SsEF-1alpha from the strain MT3 is cloned, sequenced and expressed in Escherichia coli, the vectors pGEM-3Z and pET22 are used
P35021, -
expression in Escherichia coli
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expression in Escherichia coli BL21
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expression in Escherichia coli using the pT7-7 expression vector
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expression in Escherichia coli; the SsEF-1alpha gene is cloned into the pT7-7 vector for expression in Escherichia coli BL21DE3 cells, in addition two truncated forms of SsEF-1alpha are constructed, encoding the domain G and the domains G+M
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for expression in Escherichia coli cells
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recombinant SsEF-1alpha and its nucleotide-free form are prepared by using an Escherichia coli expression system
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expression in Escherichia coli BL21
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EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
after treatment with 50 microM Zearalenone, abscisic acid, 10 microM paclobutrazole, 10 microM aminoethoxyvinylglycine, 10 microM gibberellin, 10 microM 1-N-naphthylphthalamic acid, 10 microM brassinolide, 10 microM methyl jasmonate, 45 mM nitrate and 90 mM sucrose, osmotic stress; decreased gene expression of A1, A2, A3 and A4 gene induced by abiotic stresses like salt, touch, elevated CO2; decreased gene expression of A1 and A4 gene after powdery mildew treatment; decreased gene expression of A2 with 24-h dark treatment or blue light stimulus
P13905
2fold increased gene expression level of A2 and A4 after treatment with ethylene, cytokinin, gibberellin; after interaction with mycorrhiza or nematode and treatment with 10 microM cycloheximide, 0.1 mM isoxaben, 6-benzyl adenine, 10 microM daminozide, 10 microM prohexadione, 10 microM ibuprofen, sucrose, 4C cold stress; increased A2 gene expression in response to vernalization; increased gene expression of A1, A2, A3 and A4 in response to light; stimulation of gene expression of A1 and A2 gene by Phytophthora infestans
P13905
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
P269S
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variant is expressed to a high level in Escherichia coli. The variant functions as effectively as the respective wild-type factor in ternary complex formation using Escherichia coli Phe-tRNAPhe and Cys-tRNACys. The variant is also active in A-site binding and in vitro translation assay with Escherichia coli Phe-tRNAPhe
A421(insGly)G422
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mutation causes cold-sensitivity in the organism. No GTPase activity below 10C and reduced activity at all temperatures up to 45C, as compared to wild-type enzyme
D138N
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mutant with decreased affinity for GDP and increased affinity for XDP
D409E
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mutation causes cold-sensitivity in the organism. No GTPase activity below 10C and reduced activity at all temperatures up to 45C, as compared to wild-type enzyme
D50G
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mutation reveals twofold reduction of growth rate at 30C
D80N
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mutant with decreased affinity for GTP and increased GTPase activity
E571K
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site-directed mutagenesis, the mutation in the 30S binding domain IF2-G3 disrupts hydrogen bonding between subdomains G2 and G3, so that IF2 acquires a GDP-like conformation and is no longer responsive to GTP binding. The mutant has a 6.5fold reduced affinity for the 30S subunit and is completely inactive in ribosome-dependent GTP hydrolysis. The IF2 E571K mutant is active in 30S initiation complex and initiation dipeptide formation, and supports faithful mRNA translation
G28D
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mutation reveals slightly reduced growth rate at 30C
G83A
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mutation slows down the association of the ternary complex EF-Tu/GTP/aminoacyltRNA with the ribosome and abolishes the ribosome-induced GTPase activity of EF-Tu
G83A/G94A
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mutation slows down the association of the ternary complex EF-Tu/GTP/aminoacyltRNA with the ribosome and abolishes the ribosome-induced GTPase activity of EF-Tu
G94A
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mutation strongly impairs the conformational change of EF-Tu from the GTP-bound to the GDP-bound form and decelerates the dissociation of EF-Tu/GDP from the ribosome
H448S
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site-directed mutagenesis, a dominant-lethal substitution, the expression of the mutant causes a rapid growth arrest and a reduction in the number of viable cells by 3 or 4 orders of magnitude within 20-30 min after induction
H448S/E571K
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site-directed mutagenesis, induction of the GTPase-deficient double mutant affects neither the growth of the cells nor the viable counts demonstrating that the E571K mutation is capable of suppressing lethality of the dominant-lethal H448S substitution
Q290L
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3-5fold more active in polymerization than wild-type Escherichia coli EF-Tu, 10fold increase in GTPase activity compared to wild-type enzyme
R40D
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mutation reveals reduced growth rate at 30C
R45D
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mutation reveals reduced growth rate at 30C
R45L
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mutation reveals reduced growth rate
R65D
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lethal mutation
R69D
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mutation reveals reduced growth rate at 30C
R69L
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mutation reveals reduced growth rate
R69L/R71L
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mutation reveals reduced growth rate
S221P
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variant is poorly expressed and the majority of molecules fail to fold into an active conformation. The variant functions as effectively as the respective wild-type factor in ternary complex formation using Escherichia coli Phe-tRNAPhe and Cys-tRNACys. The variant is also active in A-site binding and in vitro translation assay with Escherichia coli Phe-tRNAPhe
S69P
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mutation reveals twofold reduction of growth rate at 30C
V12A
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mutation reveals slightly reduced growth rate at 30C
D138N
Escherichia coli HW110
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mutant with decreased affinity for GDP and increased affinity for XDP
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D80N
Escherichia coli N4830
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mutant with decreased affinity for GTP and increased GTPase activity
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E424K
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random mutagenesis, the IF2-G3 domain mutant shows a reduced affinity for the 30S ribosomal subunit, the mutant shows complete loss of GTPase activity
G378C
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more stable binding within the 70S initiation complex of Bst-IF2*GDP
G420E
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random mutagenesis, the mutant shows a reduced affinity for both ribosomal subunits
S387P
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random mutagenesis, the mutant shows a reduced affinity for the 30S ribosomal subunit
A208V
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suppressor mutation to rescue growth defect associated with N135D mutation
A219T
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suppressor mutation to rescue growth defect associated with N135D mutation, as single mutant slow-growth phenotype
A382V
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suppressor mutation to rescue growth defect associated with N135D mutation
E569A
P32501
lethal mutation
E569D
P32501
lethal mutation, reduced binding to subunit gamma of eIF2
E569K
P32501
lethal mutation, reduced binding to subunit gamma of eIF2
E569Q
P32501
lethal mutation, reduced binding to subunit gamma of eIF2
H480I
P39730
impair of GTP hydrolysis and yeast cell growth
H480I/A709V
P39730
double mutant, faster yeast cell growth
H480I/F643R
P39730
double mutant, suppression of H480I-mutant mediated slow yeast cell growth
H480I/G642F
P39730
double mutant, suppression of H480I-mutant mediated slow yeast cell growth
H480I/I634G
P39730
double mutant, suppression of H480I-mutant mediated slow yeast cell growth
H480I/V637A
P39730
double mutant, suppression of H480I-mutant mediated slow yeast cell growth
H480I/V637G
P39730
double mutant, suppression of H480I-mutant mediated slow yeast cell growth
N135A
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growth defect
N135D
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slow-growth phenotype, imparied Met-tRNA binding to eIF2
N135K
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growth defect, recessive lethal mutation
S576N
P32501
slow-growing, cold sensitivity, defect on protein interaction
T439A
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reduced ribosomal subunit joining
T439A
P39730
slow-growth phenotype
T439A/F643R
P39730
double mutant, suppression of T439A-mutant mediated slow yeast cell growth
T439A/V637G
P39730
double mutant, suppression of T439A-mutant mediated slow yeast cell growth
T552I
P32501
slow-growing, cold sensitivity, defect on protein interaction
W699A
P32501
lethal mutation, weakens binding to subunit beta and gamma of eIF2, prevents nucleotide exchange
A208V
Saccharomyces cerevisiae J293
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suppressor mutation to rescue growth defect associated with N135D mutation
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A382V
Saccharomyces cerevisiae J293
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suppressor mutation to rescue growth defect associated with N135D mutation
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N135A
Saccharomyces cerevisiae J293
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growth defect
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N135D
Saccharomyces cerevisiae J293
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slow-growth phenotype, imparied Met-tRNA binding to eIF2
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N135K
Saccharomyces cerevisiae J293
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growth defect, recessive lethal mutation
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H480I
Saccharomyces cerevisiae NOY891
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impair of GTP hydrolysis and yeast cell growth
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H480I/A709V
Saccharomyces cerevisiae NOY891
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double mutant, faster yeast cell growth
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H480I/F643R
Saccharomyces cerevisiae NOY891
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double mutant, suppression of H480I-mutant mediated slow yeast cell growth
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H480I/I634G
Saccharomyces cerevisiae NOY891
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double mutant, suppression of H480I-mutant mediated slow yeast cell growth
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T439A
Saccharomyces cerevisiae NOY891
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slow-growth phenotype
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A26G
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converts the guanine nucleotide binding consensus sequences A-X-X-X-X-G-K-[T,S] of the elongation factor EF-2 into the corresponding G-X-X-X-X-G-K-[T,S] motif which is present in all the other GTP-binding proteins. In the mutant, the rate of poly(U)-directed poly(Phe) synthesis and the ribosome-dependent GTPase activity of A26GSsEF-2 are decreased. A26G substitution enhances the catalytic efficiency of the intrinsic SsEF-2 GTPase triggered by ethylene glycol and decreases the affinity for GDP
D60A
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1.4fold slower hydrolysis of GTP
F236P
-
kcat/Km is 63% compared to wild-type value
G13A
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compared to wild-type enzyme the mutant shows a reduced rate of Phe polymerization and a reduced intrinsic GTPase activity that is stimulated by high concentrations of NaCl. Mutant enzyme shows an increased affinity for GTP and GDP. The temperature inducing a 50% denaturation of the mutant enzyme is 5C lower than that of the wild-type enzyme
G235P
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complete loss of GTP hydrolyzing activity
N189P
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complete loss of GTP hydrolyzing activity
Ss(G)EF-1alpha
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truncated form of SsEF-1alpha
Ss(GM)EF-1alpha
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truncated form of SsEF-1alpha
T193N
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complete loss of GTP hydrolyzing activity
T213V
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kcat is 45% compared to the wild-type value
F236P
-
kcat/Km is 63% compared to wild-type value
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N189P
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complete loss of GTP hydrolyzing activity
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T193N
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complete loss of GTP hydrolyzing activity
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A375T
Thermus thermophilus JC469
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resistance to kirromycin, abolished streptomycin resistance of mutants of ribosomal protein S12
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additional information
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Snowy cotyledon 1 mutant contains a mutation in a gene encoding the chloroplast elongation factor G, leading to an amino acid exchange within the predicted 70S ribosome-binding domain. The mutation results in a delay in the onset of germination. At this early developmental stage embryos still contain undifferntiated proplastids, whose proper function seems necessary for seed germination. In light-gropwn sco1 seedlings the greening of cotyledons is severely impaired, whereas the following true leaves develop normally as in wild-type plants
H84A
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reduces the rate constant of GTP hydrolysis more than 1000000fold, the preceding steps of ternary complex binding to the ribosome, codon recognition and the GTPase activation step are affected only slightly. The catalytic role of His84 in elongation factor Tu is to stabilize the transition state of GTP hydrolysis by hydrogen bonding to the attacking water molecule or, possibly, the gamma-phosphate group of GTP
additional information
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used as a hybrid sytem with Thermus thermophilus ribsomal protein L11
additional information
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two EF-G cysteine mutants 58C and 196C react efficiently with 2',7'-difluorofluorescein maleimide, whereas the cysteine-free protein is unreactive
additional information
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GTPase null mutant E424K of Bacillus stearothermophilus can replace in vivo wild-type IF2 allowing the Escherichia coli infB null mutant to grow with almost wild-type duplication times
H301Y
-
site-directed mutagenesis, GTPase-deficient mutant
additional information
-
GTPase null mutant E424K of Bacillus stearothermophilus can replace in vivo wild-type IF2 allowing the Escherichia coli infB null mutant to grow with almost wild-type duplication times
L568A
P32501
cold sensitivity, defect on protein interaction
additional information
P32501
inherited mutations cause fatal brain disorder: childhood ataxia with central nervous system hypomyelination, leukoencephalopathy with vanishing white matter, eIF2B-related disorders; mutations in subunit beta destabilize interaction between eIF2, eIF1A, eIF3 and eIF5
G235S
-
partial loss of GTP hydrolyzing activity
additional information
-
construction of truncated forms corresponding to the putative domains G+M, and domain G. Neither truncated form is able to sustain poly(Phe) synthesis but they are able to bind guanine nucleotides with an affinity much higher with respect to that of the intact factor. Kinetic data are not changed by the truncation, but both forms are less thermostable than the intact factor and both are no more sensitive to the stimulatory effect of elongation factor 1beta
additional information
P35021, -
comparison of sequence with that of Sulfolobus solfataricus strain MT4 shows only one amino acid change, i.e. I15V. The difference is in the first guanine nucleotide binding consensus sequence G13HIDHGK and is responsible for a increased efficiency in protein synthesis, which is accompanied by an reduced affinity for both guanosine diphosphate (GDP) and guanosine triphosphate (GTP), and an decreased efficiency in the intrinsic GTPase activity. The exchange has only very marginal effects on the thermal properties of the enzyme; comparison of sequence with that of Sulfolobus solfataricus strain MT4 shows only one amino acid change, i.e. V15I. The difference is in the first guanine nucleotide binding consensus sequence G13HIDHGK and is responsible for a reduced efficiency in protein synthesis, which is accompanied by an increased affinity for both guanosine diphosphate (GDP) and guanosine triphosphate (GTP), and an increased efficiency in the intrinsic GTPase activity. The exchange has only very marginal effects on the thermal properties of the enzyme
additional information
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the N-terminal deletion mutant displays a similar Km value as the wild-type enzyme whereas the substrate kcat is 24fold increased
G235P
-
complete loss of GTP hydrolyzing activity
-
additional information
-
the N-terminal deletion mutant displays a similar Km value as the wild-type enzyme whereas the substrate kcat is 24fold increased
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T213V
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kcat is 45% compared to the wild-type value
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additional information
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mutant of elongation factor G containing the effector loop from Thermus aquaticus EF-Tu has markedly decreased GTPase activity and does not catalyze translocation. The loops are not functionally interchangeable since the factors interact with different states of the ribosome
A375T
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resistance to kirromycin, abolished streptomycin resistance of mutants of ribosomal protein S12
additional information
-
mutant of elongation factor G containing the effector loop from Thermus aquaticus EF-Tu has markedly decreased GTPase activity and does not catalyze translocation. The loops are not functionally interchangeable since the factors interact with different states of the ribosome
additional information
-
used as a hybrid sytem with Escherichia coli ribsomal proteins L10, L11, L12
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
denaturation by urea and guanidine hydrochloride shows a cooperative unfolding process with no intermediate species. Chemical unfolding by urea and guanidine hydrochloride is fully reversible for both enzyme-GDP complex and nucleotide-free enzyme. Both forms exhibit remarkable stability against urea, but not against guanidine hydrochloride
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APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
polypeptide chain synthesis
medicine
-
chaperon-activity; protein biosynthesis
medicine
-
elongation factor Tu may have in vivo role in tetracycline inhibition of protein synthesis
medicine
Escherichia coli HW110
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polypeptide chain synthesis
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medicine
Escherichia coli MRE600
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protein biosynthesis; protein biosynthesis
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medicine
-
termination of ribosome-dependent protein synthesis
medicine
-
protein biosynthesis
medicine
Pseudomonas aeruginosa 8830
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protein biosynthesis
-
medicine
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protein biosynthesis
medicine
-
polyphenylalanine synthesis
medicine
Saccharomyces carlsbergensis EF-3
-
polyphenylalanine synthesis
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medicine
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chaperon-activity; protein biosynthesis
medicine
-
polyphenylalanine synthesis
medicine
Schizosaccharomyces pombe EF-3
-
polyphenylalanine synthesis
-
medicine
-
polypeptide chain synthesis
medicine
-
chaperon-activity; protein biosynthesis
medicine
-
polypeptide chain synthesis