Information on EC 2.7.7.72 - CCA tRNA nucleotidyltransferase

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

EC NUMBER
COMMENTARY hide
2.7.7.72
-
RECOMMENDED NAME
GeneOntology No.
CCA tRNA nucleotidyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
a tRNA precursor + 2 CTP + ATP = a tRNA with a 3' CCA end + 3 diphosphate
show the reaction diagram
a tRNA precursor + CTP = a tRNA with a 3' cytidine end + diphosphate
show the reaction diagram
-
-
-
-
a tRNA with a 3' CC end + ATP = a tRNA with a 3' CCA end + diphosphate
show the reaction diagram
-
-
-
-
a tRNA with a 3' cytidine + CTP = a tRNA with a 3' CC end + diphosphate
show the reaction diagram
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
CTP,CTP,ATP:tRNA cytidylyl,cytidylyl,adenylyltransferase
The acylation of all tRNAs with an amino acid occurs at the terminal ribose of a 3' CCA sequence. The CCA sequence is added to the tRNA precursor by stepwise nucleotide addition performed by a single enzyme that is ubiquitous in all living organisms. Although the enzyme has the option of releasing the product after each addition, it prefers to stay bound to the product and proceed with the next addition [5].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
Escherichia coli B / ATCC 11303
-
-
-
Manually annotated by BRENDA team
strain ATCC 51573
-
-
Manually annotated by BRENDA team
no activity in Saccharomyces cerevisiae
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
a tRNA precursor + 2 CTP + ATP
a tRNA with a 3' CCA end + 3 diphosphate
show the reaction diagram
a tRNA precursor + 2 CTP + ATP
a tRNA with a 3'CCA end + 3 diphosphate
show the reaction diagram
a tRNA precursor + CTP
a tRNA with a 3' cytidine end + diphosphate
show the reaction diagram
a tRNA with a 3' CC end + ATP
a tRNA with a 3' CCA end + diphosphate
show the reaction diagram
a tRNA with a 3' CCA end + 3 diphosphate
a tRNA precursor + 2 CTP + ATP
show the reaction diagram
a tRNA with a 3' cytidine + CTP
a tRNA with a 3' CC end + diphosphate
show the reaction diagram
a tRNA(Ala) precursor + 2 CTP + ATP
a tRNA(Ala) with a 3' CCA end + 3 diphosphate
show the reaction diagram
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synthesizes poly(C) when incubated with CTP alone, but switches to synthesize CCA when incubated with both CTP and ATP. The enzyme also exhibits a processing activity that removes nucleotides in the 3' to 5' direction to as far as position 74
-
-
?
ATP + tRNA-C-C
tRNA-C-C-A + diphosphate
show the reaction diagram
CTP + tRNA-C
tRNA-C-C + diphosphate
show the reaction diagram
-
-
-
-
?
deoxynucleoside triphosphate + DNAn
diphosphate + DNAn+1
show the reaction diagram
tRNA(Phe) precursor + 2 CTP + ATP
tRNA(Phe) with a 3' CCA end + 3 diphosphate
show the reaction diagram
-
the 48 kDa monomer forms a stable salt-resistant dimer in solution. Further dimerization of the dimeric enzyme to form a tetramer is induced by the binding of two tRNA molecules. The formation of a tetramer with only two bound tRNA molecules leads to the suggestion that one pair of active sites may be specific for adding two C bases, which results in scrunching of the primer strand. An adjacent second pair of active sites may be specific for adding A after addition of two C bases which makes the 30 terminus long enough to reach the second pair of active sites
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-
?
tRNAAsp with a 3' CC end + ATP
tRNAAsp with a 3' CCA end + diphosphate
show the reaction diagram
tRNAAsp with a 3' cytidine + CTP
tRNAAsp with a 3' CC end + diphosphate
show the reaction diagram
tRNACys + 2 CTP + ATP
tRNACys with 3'-CCA end + 3 diphosphate
show the reaction diagram
-
insertional editing of substrate is not required for addition of the CCA sequence by CCase
-
-
?
tRNAX1 + ATP + 2 CTP
tRNAXCCA + 3 diphosphate
show the reaction diagram
yeast tRNAPhe + 2 CTP + ATP
yeast tRNAPhe with 3'-CCA end + 3 diphosphate
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
a tRNA precursor + 2 CTP + ATP
a tRNA with a 3' CCA end + 3 diphosphate
show the reaction diagram
a tRNA with a 3' CCA end + 3 diphosphate
a tRNA precursor + 2 CTP + ATP
show the reaction diagram
deoxynucleoside triphosphate + DNAn
diphosphate + DNAn+1
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mn2+
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while Mn2+ is incompetent for diphosphorolysis of tRNA-A76, it promotes a more rapid forward synthesis of tRNA-A76 than even the Mg2+ ion
Zn2+
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it is inert for diphosphorolysis of tRNAA76, but it is active in the forward synthesis of this tRNA
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
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inhibition of AMP incorporating activity only, the CMP incorporating activity is much less sensitive
1,7-phenanthroline
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slight inhibition
2,2,2-Terpyridyl
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inhibition of AMP incorporating activity only, the CMP incorporating activity is much less sensitive
2,2-dipyridyl
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at high concentrations, inhibition of AMP incorporating activity only, the CMP incorporating activity is much less sensitive
bathophenanthroline
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inhibition of AMP incorporating activity only, the CMP incorporating activity is much less sensitive
ethylnitrosourea
Neocuproine
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slight inhibition
additional information
-
no inhibition by EDTA
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
protein Hfq
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.008 - 0.119
a tRNA precursor
-
0.054 - 12.5
ATP
0.01 - 0.65
CTP
0.5 - 1
diphosphate
additional information
ATP
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.09 - 0.11
a tRNA precursor
-
0.0013 - 2.72
ATP
0.0012 - 3.8
CTP
1.4 - 3.1
diphosphate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.07 - 1.23
a tRNA precursor
-
0.005 - 0.7
ATP
0.0033 - 0.09
CTP
1.4 - 5.2
diphosphate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
67.1
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purified enzyme, pH not specified in the publication, 37C, pooled column fraction
280
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purified enzyme, pH not specified in the publication, 37C, column peak fraction
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 10
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the enzyme exhibits substantial activity over a broad range from pH 7 to 10
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
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weak activity at 50C, no activity at 37C
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.85
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both AMP- and CMP-incorporating activities, isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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optimal growth condition of the organismat 83C
Manually annotated by BRENDA team
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TRNT1 protein levels are 10fold lower in muscle than in fibroblasts
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
51500
-
1 * 51500, SDS-PAGE
53000
-
gel filtration
98000
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dimer, gel filtration
242000
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tetrameric enzyme with two bound tRNA(Phe)s, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 48000, SDS-PAGE
dimer
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2 * 48000, the 48 kDa monomer forms a stable salt-resistant dimer in solution. Further dimerization of the dimeric enzyme to form a tetramer is induced by the binding of two tRNA molecules
monomer
tetramer
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2 * 48000, the 48 kDa monomer forms a stable salt-resistant dimer in solution. Further dimerization of the dimeric enzyme to form a tetramer is induced by the binding of two tRNA molecules
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cocrystal structures of the enzyme complexed with both a tRNA mimic and nucleoside triphosphates under catalytically active conditions. The structures suggest that adenosine 5'-monophosphate is incorporated onto the A76 position of the tRNA via a carboxylate-assisted, one-metal-ion mechanism with aspartate 110 functioning as a general base. The discrimination against incorporation of cytidine 5'-triphosphate at position 76 arises from improper placement of the a phosphate of the incoming CTP, which results from the interaction of C with arginine 224 and prevents the nucleophilic attack by the 3' hydroxyl group of cytidine75
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cocrystallisation of enzyme complexes with nine distinct tRNA minihelices. All of the binary and ternary complex crystals belong to the space group P4(3)2(1)2, and contain one complex molecule in the asymmetric unit. Crystal structures are solved at 2.5 to 3.05 A resolutions by molecular replacement
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crystal structure analysis
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crystal structures of the CCA-adding enzyme, and its complexes with CTP and ATP at 2.0, 2.0 and 2.7 A resolutions, respectively
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hanging-drop vapour diffusion method at 4C, crystal structure of the enzyme in complex with tRNA
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in complex with a human MenBeta minihelix. The unstable minihelix is bound between the enzymes catalytic center, comprised of the head and neck domains, and its tail domain. The minihelix perfectly mimics full-length tRNA with its acceptor and TPsiC stems folding into a continuous A-type RNA helix. The TPsiC loop is in the same conformation as in full-length tRNA
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crystal structure analysis
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modeling of a loop sequence inserted into the structure of human CCA-adding enzyme based on pdb-entry 1OU5. The conserved loop residue R105 forms a salt bridge to the first residue E164 of the amino acid template EDxxR in motif D
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purified recombinant monomeric native enzyme or enzyme in complex with thimerosal, sitting drop vapor diffusion method, mixing of 0.003-0.005 ml of protein solution with 0.001 ml of reservoir solution containing 100 mM sodium citrate, pH 5.6, 2.2 M ammonium sulfate, and equilibration against 0.4 ml of reservoir solution, 3 days to 3 weeks, for ligand bound enzyme crystals soaking in reservoir solution containing 0.5 mM thimerosal for 45 min, 18C, X-ray diffraction structure determination and analysis at 3.4-3.7 A resolution, single isomorphous replacement
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crystal structure analysis
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme 11800fold by ammonium sulfate fractionation, anion exchange and hydroxylapatite chromatography, and tRNA affinity chromatography
-
purified by a combination of ammonium sulfate fractionation, gel filtration, and hydrophobic interaction chromatography (NTSFII); purified by a combination of anion-exchange chromatography and hydrophobic interaction chromatography
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recombinant C-terminally His-tagged enzyme from Escherichia coli
recombinant His-tagged HD domain from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography
recombinant N-terminally His-tagged mitochondrial CCase from Escherichia coli strain BL21 CodonPlus (DE3)-RP by nickel affinity chromatography and gel filtration to over 95% purity
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
expressed in Escherichia coli BL21(DE3)/pLysS Rosetta cells
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expression in Escherichia coli
mitochondrial CCase, expression as N-terminally His-tagged enzyme in Escherichia coli strain BL21 CodonPlus (DE3)-RP
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overexpression in Escherichia coli
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overexpression of C-terminally His-tagged enzyme in Escherichia coli
recombinant expression of the isolated His-tagged HD domain in Escherichia coli strain BL21 (DE3)
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E189F
-
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the temperature-sensitive phenotype of mutant E189F results from a reduced ability to incorporate AMP and CMP into tRNAs. This defect can be compensated for by a second-site suppressor converting residue arginine 64 to tryptophan. The R64W substitution does not alter the structure or thermal stability of the enzyme dramatically but restores catalytic activity of mutant E189F in vitro and suppresses the temperature-sensitive phenotype in vivo. Residues R64 and E189 do not interact directly, residue E189 has a role in enzyme structure and function
P295A
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kcat /Km for C74 addition is 28% of wild-type value, kcat /Km for C75 addition is 33% of wild-type value, kcat /Km for A76 addition is 1% of wild-type value
P295G
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kcat /Km for C74 addition is 94% of wild-type value, kcat /Km for C75 addition is 15% of wild-type value, kcat /Km for A76 addition is 129% of wild-type value
P295S
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kcat /Km for C74 addition is 3% of wild-type value, kcat /Km for C75 addition is 1% of wild-type value, kcat /Km for A76 addition is 1% of wild-type value
P295T
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kcat /Km for C74 addition is 36% of wild-type value, kcat /Km for C75 addition is 67% of wild-type value, kcat /Km for A76 addition is 92% of wild-type value
.R190I
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homozygous mutation identified in a patient with sideroblastic anemia with B-cell immunodeficiency, periodic fevers, and developmental delay
D139A
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mutant shows a strong reduction in the addition of the terminal A position
G143A
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similar to wild-type, mutant catalyzes the addition of the complete CCA sequence
L166S
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and T154I, compound heterozygous mutation identified in a patient with sideroblastic anemia with B-cell immunodeficiency, periodic fevers, and developmental delay
R153A
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similar to wild-type, mutant catalyzes the addition of the complete CCA sequence
T154I
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and L166S, compound heterozygous mutation identified in a patient with sideroblastic anemia with B-cell immunodeficiency, periodic fevers, and developmental delay
A75C
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as compared to wild-type enzyme the Km increases 2.7-fold from A addition to C addition, while the kcat decreased 14fold. The overall kcat/Km ratio of C addition to A addition is 0.03
D106A
-
CTP-adding activity is 90% of wild-type level, ATP-adding activities is 14% of wild-type level; mutation has no effect on CCA-adding activity
D124A
-
mutation has no effect on CCA-adding activity
D143A
-
mutation has no effect on CCA-adding activity
D215A
-
CTP-adding activity is 16% of wild-type level, ATP-adding activities is 13% of wild-type level; mutation has no effect on CCA-adding activity
D218A
-
mutation has no effect on CCA-adding activity
D53A
-
no activity
D53A/D55A
-
no activity
D55A
-
no activity
D55E
-
no activity
delC135
-
no activity
delC30
-
no activity
E104A
-
mutation has no effect on CCA-adding activity
E114A
-
mutation has no effect on CCA-adding activity
E139A
-
mutation has no effect on CCA-adding activity
E144A
-
mutation has no effect on CCA-adding activity
E161A
-
mutation has no effect on CCA-adding activity
E173A
-
CTP-adding activity is 19% of wild-type level, ATP-adding activities is 7% of wild-type level; mutation has no effect on CCA-adding activity
E209A
-
mutation has no effect on CCA-adding activity
E92F
-
the mutant is nearly inactive for CCA addition, although tRNA binding is apparently normal
G156D
-
neither CTP- nor ATP-adding activity is significantly affected
G166R
-
C74-adding is 7% of wild-type activity, C75-adding is 8% of wild-type activity, A76-adding is 21% of wild-type activity
G169R
-
C74-adding is 5% of wild-type activity, C75-adding is 2% of wild-type activity, A76-adding is 22% of wild-type activity
H129A
-
C74-adding is 22% of wild-type activity, C75-adding is 28% of wild-type activity, A76-adding is 87% of wild-type activity
H93V
-
the mutant adds C74 and C75 but fails to add A76
H93V/Y95V
-
the mutant enzyme is completely inactive for CCA addition
K149A
-
no C74-adding activity, C75-adding is 1% of wild-type activity, A76-adding is 1% of wild-type activity
K153A
-
C74-adding is 14% of wild-type activity, C75-adding is 4% of wild-type activity, A76-adding is 3% of wild-type activity
R125A
-
C74-adding is 30% of wild-type activity, C75-adding is 10% of wild-type activity, A76-adding is 5% of wild-type activity
V34C
-
as compared to wild-type enzyme the Km increases 1.4fold from A addition to C addition, while the kcat decreases 18fold. The overall kcat/Km ratio of C addition to A addition is 0.04
Y158A
-
C74-adding is 9% of wild-type activity, C75-adding is 15% of wild-type activity, A76-adding is 7% of wild-type activity
Y90E
-
the mutant is nearly wild type for CCA addition despite replacement of a conserved bulky hydrophobic residue by a carboxylate
Y90E/E92F
-
the mutant enzyme is completely inactive for CCA addition
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
medicine
Show AA Sequence (9696 entries)
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