Information on EC 3.6.4.9 - chaperonin ATPase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.6.4.9
-
RECOMMENDED NAME
GeneOntology No.
chaperonin ATPase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + H2O = ADP + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
hydrolysis of phosphonic ester
-
-
-
-
hydrolysis of phosphoric ester
SYSTEMATIC NAME
IUBMB Comments
ATP phosphohydrolase (polypeptide-unfolding)
Multisubunit proteins with 2x7 (Type I, in most cells) or 2x8 (Type II, in Archaea) ATP-binding sites involved in maintaining an unfolded polypeptide structure before folding or entry into mitochondria and chloroplasts. Molecular masses of subunits ranges from 10-90 kDa. They are a subclass of molecular chaperones that are related to EC 3.6.4.8 (proteasome ATPase).
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain K1
-
-
Manually annotated by BRENDA team
anoxygenic photosynthetic purple sulfur bacterium
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
thermophilic strain MS, highly homologous to Bacillus stearothermophilus
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
used in complex with gp23 of bacteriophage T4 and GroEL of Escherichia coli; used in complex with gp31 of bacteriophage T4 and GroEL of Escherichia coli
-
-
Manually annotated by BRENDA team
mouse
-
-
Manually annotated by BRENDA team
gene groEL
UniProt
Manually annotated by BRENDA team
gene groEL
UniProt
Manually annotated by BRENDA team
purple non-sulfur photosynthetic bacterium
-
-
Manually annotated by BRENDA team
gram-positive, thermophilic anaerobe
-
-
Manually annotated by BRENDA team
strain 7, acidothermophilic archaeon
-
-
Manually annotated by BRENDA team
strain 7, acidothermophilic archaeon
-
-
Manually annotated by BRENDA team
strain KOD1
-
-
Manually annotated by BRENDA team
Cpnalpha
SwissProt
Manually annotated by BRENDA team
-
-
-
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
ADP + H2O
AMP + phosphate
show the reaction diagram
AMP + H2O
adenosine + phosphate
show the reaction diagram
ATP + H2O
ADP + phosphate
show the reaction diagram
CDP + H2O
CMP + phosphate
show the reaction diagram
-
-
-
?
CTP + H2O
CDP + phosphate
show the reaction diagram
GDP + H2O
GMP + phosphate
show the reaction diagram
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
IDP + H2O
IMP + phosphate
show the reaction diagram
-
-
-
?
ITP + H2O
IDP + phosphate
show the reaction diagram
-
-
-
?
UDP + H2O
UMP + phosphate
show the reaction diagram
-
-
-
?
UTP + H2O
UDP + phosphate
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
ADP + H2O
AMP + phosphate
show the reaction diagram
ATP + H2O
ADP + phosphate
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
GroES
-
additional information
-
GroES encapsulates substrate protein
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cs+
-
1 mM, 81% of K+ activation
Cu2+
-
the enzyme is less active, when Ca2+, Cu2+, Co2+, and Ni2+ are used ar the same concentration instead of Mn2+
KCl
-
50-500 mM, stimulates ATPase activity. ATPase activity decreases significantly at higher KCl concentrations (1-2 M)
Li+
-
1 mM, 74% of K+ activation
Na+
-
1 mM, 87% of K+ activation
NaCl
-
50-500 mM, stimulates ATPase activity. The enzyme maintains full ATPase activity up to 2 M
NH4Cl
-
50-500 mM, stimulate ATPase activity. The enzyme maintains full ATPase activity up to 2 M
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-Chloro-7-nitrobenzo-2-oxa-1,3-diazole
-
0.1 mM, 31% inhibition
AMP
-
when AMP concentration is 8 times that of ATP, AMP inhibits the ATPase activity to 48%
AMP-PNP
-
2 mM, 90% inhibition
arsenic
-
arsenic inhibits TRiC function, required for folding of actin, tubulin, and other proteins post-synthesis
ATP-gammaS
-
complete inhibition of ATPase activity
beryllium fluoride
-
BeFx
Cpn10
-
Dicyclohexylcarbodiimide
-
0.1 mM, 43% inhibition
EC3016
-
inhibits ATPase activity of mutant I493C
GroES
-
GroESx
-
approx. 40% inhibition above a molar ratio of 1 to 1
-
Guanidinium chloride
-
20% inhibition with 150 mM guanidinium chloride in the presence of 100 mM KCl, 80% inhibition with 1 mM KCl
KCl
-
50-500 mM, stimulates ATPase activity. ATPase activity decreases significantly at higher KCl concentrations (1-2 M)
Mg2+
-
inhibits the phosphorylation of enzyme during hydrolysis
MgADP
-
50% inhibition at 0.205 mM
Mn2+
-
25% reversible inhibition of Mg-ATPase activity
MnADP
-
50% inhibition at 0.036 mM
N-ethylmaleimide
-
0.1 mM, 32% inhibition
N-[4-(4-amino-1-tert-butyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl]cyclopentanecarboxamide
-
specifically blocks ATP binding and hydrolysis
Na2SO3
-
200 mM, 86% inhibition
SO42-
-
mildly inhibits the K+-dependent ATP hydrolysis
yeast mitochondrial chaperonin hsp10
-
additional information
-
physiological ADP concentration suppresses formation of symmetric complexes
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ammonium sulfate
-
optimal functionality of MmGroEL/GroES and its ability to encapsulate larger proteins, such as malate dehydrogenase, requires the presence of ammonium sulfate in vitro
ATP
-
ATP is an allosteric ligand for GroEL, its binding promoting both cooperative (intra-ring) and anti-cooperative (inter-ring) actions. ATP serves as a substrate, undergoing hydrolysis during the reaction cycle to promote a unidirectional advance of the machine
Cpn10
-
denatured beta actin
-
4fold increase in ATPase activity at 30C
-
denatured beta tubulin
-
-
-
GroES
-
K+
-
enhances the phosphorylation of enzyme during hydrolysis
N-ethylmaleimide
-
2fold increase in activity after modification of GroEL with 2 mM N-ethylmaleimide
NaHCO3
-
200 mM, 2.15fold increase in activity
phosducin-like protein 2
-
i.e. PLP2, stimulatory cofactor. 30fold stimulation of actin folding
-
unfolded substrate protein
-
chaperonin machine enhances hemicycle time and mean residence time set by the rate of ATP hydrolysis by the cis ring
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.007 - 10.04
ATP
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000667 - 2.21
ATP
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.006
ATPase activity at 40C
0.0193
-
at 37C
0.031
-
alpha chaperonin subunit
0.751
-
at 37C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.8
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 6
-
pH 4.0: about 45% of maximal activity, pH 6.0: about 65% of maximal activity
4 - 11
-
pH 4.0: about 65% of maximal activity, pH 11.0: about 60% of maximal activity
6 - 9
-
almost no activity at pH 6.0, approx. 90% of maximal activity at pH 9.0
7 - 9
-
pH 6.5: about 30% of maximal activity, pH 7.0: about 90% of maximal activity, pH 9.0: about 60% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
assay at
22 - 37
-
release/reactivation of rhodanese from/by GroEL
23
-
assay at
49
-
in the presence of 2 mM K+ and Mg2+
60 - 80
-
authentic and recombinant Cpn60
100
-
while complexed to heat inactivated lysozyme, Pf Cpn shows increased optimal temperature for ATPase activity from 90C to 100C
additional information
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 37
-
and above, the release/reactivation of rhodanese from/by GroEL is minimal at 4C, but is optimal between 22C and 37C
25 - 80
-
His6-tagged chaperonin Cpn60
25 - 60
-
approx. 50% of maximal activity at 30C, approx. 90% of maximal activity at 55C, almost no activity at 60C
25 - 64
-
ATPase activity gradually increases between 25C and 52C, strong decrease above
25 - 70
30 - 65
35 - 85
CTT complex, approx. 50% of maximal activity at 70C, approx. 80% of maximal activity at 75C, in the presence of 2.9 N KCl
40 - 90
-
approx. 15% of maximal activity at 40C; approx. 40% of maximal activity at 40C, approx. 10% of maximal activity at 90C
40 - 70
-
0.15 mM released phosphate at 40C, 0.35 mM released phosphate at 70C
60 - 90
-
no or very little activity below 40C
70 - 95
-
70C: about 50% of maximal activity, 95C: about 70% of maximal activity
70 - 90
70C: about 55% of maximal activity (CPnalpha), about 60% of maximal activity (CPNalphabeta)
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
predicted pI-value
5.38
sequence calculation
5.49
-
calculated from sequence
7.1
deduced from nucleotide sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
CPN10 is a heat shock protein and shows increased intracellular concentration at an elevated temperature and in axenically cultivated amastigotes; GroES homologue
Manually annotated by BRENDA team
-
overexpression of Hsp60 does not extend the in vitro lifespan of normal human fibroblasts
Manually annotated by BRENDA team
-
knock-down of Hsp60 causes their growth arrest
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain UTI89 / UPEC)
Thermococcus sp. (strain JCM 11816 / KS-1)
Thermococcus sp. (strain JCM 11816 / KS-1)
Thermococcus sp. (strain JCM 11816 / KS-1)
Thermococcus sp. (strain JCM 11816 / KS-1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
56000
x * 56000, alpha subunit, + x * 56000, beta subunit, SDS-PAGE; x * 56000, alpha subunit, + x * 56000, beta subunit, SDS-PAGE
57179
x * 57179, deduced from nucleotide sequence
57259
-
x * 57259, sequence calculation, x * 60000, recombinant enzyme, SDS-PAGE
57260
-
monomer
57949
-
7 * 63000, SDS-PAGE, 7 * 57949, electrospray mass spectrometry
62000
x * 56915.4, mass spectrometry, x * 62000, recombinant His-tagged enzyme, SDS-PAGE
63000
-
7 * 63000, SDS-PAGE, 7 * 57949, electrospray mass spectrometry
66200
determined by SDS-PAGE
76200
x * 76200, immunoblot
86300
x * 86300, immunoblot
95400
x * 95400, immunoblot
450000
800000
889200
-
gel filtration
900000
-
gel filtration, recombinant Mm-cpn
920000
-
gel filtration
960000
-
-
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
the unusual dimerization of the protein leads to exposure of certain hydrophobic patches on the surface of the protein, and it is hypothesized that this might have relevance in binding to immunogenic peptides, as it does in the eukaryotic homologs
heptamer
heterooligomer
-
complex formed by two rings connected back-to-back, each composed of eight distinct subunits (CCTalpha to CCTzeta)
hexadecamer
homooligomer
-
alpha16mer; beta16mer
homotetradecamer
monomer
octamer
-
ATP-dependent subunit allows conformational change of chaperonin
oligomer
tetradecamer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
analysis of crystal structures of the GroE chaperonin from Escherichia coli in the open, ATP-bound, PDB ID 1KP8, and closed, PDB ID 1AON, states
-
analysis of the X-ray crystal structures of GroEL and GroEL-GroES complexes in absence or presence of ATP, PDB IDs 1OEL and 1SVT, and of enzyme mutant E461K interface, PDB ID 2EU1
-
crystals of GroELE461K are grown at 4C in hanging-drop vapour-diffusion experiments. The crystal structure of the mutant chaperonin GroELE461K is determined at 3.3 A and compared with other structures
-
frozen-hydrated GroEL complexes, 30A resolution, 7fold symmetry, two interring contacts per subunit, ATP turnover causes allosteric switching between the rings by altering the interring contacts
-
sitting-drop vapour-diffusion method. The well solution contains 100 mM Na-Hepes (pH 7.5), 20% (w/v) PEG 4000, 200 mM (NH4)2SO4. Concentration of protein in a drop, after mixing with an equal volume of precipitant solution, is 1520 mg/ml. Crystals appears after one week at 18C
-
hanging drop vapor diffusion method by using a reservoir solution containing 100 mM HEPES (pH 7.5), 25% (wt/vol) polyethylene glycol 3350, and 10% (vol/vol) n-propanol, 3.2 A resolution
-
2-dimensional crystals of the beta subunits of the chaperonin TF55 from Sulfolobus shibatae reconstituted into oligomers in the absence of alpha subunits. The subunits form rings with 9-fold rotational symmetry which arrange themselves in a trigonal lattice
-
electron cryo-microscopy study. Enzyme shows symmetric open and closed forms and an asymmetric form resembling a flattened GroEL-GroES bullet complex. The closed rings of the Sulfolobus chaperonin correspond to nucleotide-bound, and the open rings to nucleotide-free forms
-
electron microscopy and circular dichroism study reveal two distinct conformational states that are part of chaperonin functional cycle. The closed archaeosome complex binds ATP and forms an open complex. Upon ATP hydrolysis, the open complex dissociates into subunits. Free subunits reassemble into a two-ring structure
-
atomic force microscope imaging of enzyme without any preparation technique. Evidence for the presence of two distinct structures of the complex, which probably correspond to the open and closed conformations. Enzyme exists in solution as both single-ring and double-ring complexes
-
electron microscopy image shows a ninefold symmetrical complex about 17.5 nm in height and 16 nm in diameter, with a central cavity of 4.5 nm
-
two-dimensional crystals with p312 symmetry. Protein forms complexes of two stacked nine-subunit rings with threefold symmetry
-
visualization of enzyme by transmission electron microscopy shows a ring complex, with a diameter of the ring of 16.7 nm, and a dark cavity of 5 nm diameter present when viewed from the top; visualization of enzyme by transmission electron microscopy shows a ring complex, with a diameter of the ring of 16.7 nm, and a dark cavity of 5 nm diameter present when viewed from the top
hanging-drop, 5-10 mg/ml protein in 20 mM 4-morpholinepropanesulfonic acid, pH 7, 40 mM MgCl2 and 10% 2-propanol, 15-25% 2-propanol in reservoir, rhomboidal crystals appear overnight
-
crystal structure of the native GroEL-GroES-ADP homolog with substrate proteins in the cis-cavity, at 2.8 A resolution. Twenty-four in vivo substrate proteins within the cis-cavity are identified from the crystals
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2
-
dissociation below
681368
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35
-
half-life at 70C, 30 min, less than 5% activity retained after 90 min
43
-
GroEL interacts strongly with the enzyme rhodanese undergoing thermal unfolding at 43C. The enzyme forms a binary complex. Active rhodanese (82%) could be recovered by releasing the enzyme from GroEL after the addition of several components, e.g. ATP and the co-chaperonin GroES. The inability to recover active enzyme at 43C from the GroELrhodanese complex is not due to the disruption of the complex or aggregation of rhodanese, but rather to the partial loss of its ATPase activity and/or to the inability of rhodanese to be released from GroEL due to a conformational change
60
-
significant loss of activity
75
no loss of activity after 30 min
80 - 95
-
dissociation of alpha16mer into ATPase inactive monomers; dissociation of beta16mer into ATPase inactive monomers
80
-
50% loss of activity after 88 min; approx. 60% loss of activity after 90 min
85
-
50% loss of activity after 27 min; 50% loss of activity after 42 min
100
-
3 h, 20% loss of activity
105
-
t1/2: 141 min
110
-
t1/2: 82 min
additional information
-
while complexed to heat inactivated lysozyme, Pf Cpn shows enhanced thermostability
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
GroEL is rather insensitive to oxidants produced endogenously during metabolism, such as nitric oxide or hydrogen peroxide, but is efficiently modified and inactivated by reactive species generated by phagocytes, such as peroxynitrite and hypochlorous acid (HOCl). HOCl inactivates through the oxidation of methionine to methionine sulfoxide. In addition to the oxidation of methionine, HOCl causes the conversion of cysteine to cysteic acid and this product may account for the remainder of inactivated GroEL not recoverable through MsrB/A. HOCl produces only negligible yields of 3-chlorotyrosine. The high sensitivity of GroEL toward HOCl and ONOO suggests that this protein may be a target for bacterial killing by phagocytes
-
protein substrate binding stabilized the hexadecamer of Pf Cpn in 3 M Gdn-HCl guanidine hydrochloride
-
the enzyme complex is resistant to denaturing agents at room temperature and only pH values lower than 2 lead to dissociation
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ethanol
guanidine hydrochloride
-
protein substrate binding stabilized the hexadecamer of Pf Cpn in 3 M Gdn-HCl guanidine hydrochloride
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
22C, stable for several days in presence of up to 8 M urea
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
70C, 30 min, hydrophobic chromatography, partially purified alpha and beta subunits
-
affinity purification reveals the association of WD40 protein constitutive photomorphogenic 1 with the hetero-oligomeric TCP-1 chaperonin complex in mammalian cells
-
C6 adsorbent, Q Sepharose, 95-98% purity
-
chaperonin complexes
gel filtration
-
GroEL is purified from lysates of Escherichia coli bearing the multicopy plasmid pGroESL
-
Mono Q HR, ATP-agarose
-
more than 95% pure after SDS-PAGE
-
protein is extracted by homogenisation of deyolked embryos
-
recombinant alpha chaperonin subunit
-
recombinant alpha16mer; recombinant beta16mer
-
recombinant chaperonin GroEL from Escherichia coli
-
recombinant Cpn60-1; recombinant Cpn60-2; recombinant Cpn60-3
-
recombinant GroEL
-
recombinant GroEL wild-type and mutants from Escherichia coli strain BL21(DE3)
-
recombinant GroELx
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant Mm-cpn, Mono Q, Superdex S-300
-
Sephacryl S-300, Q-Sepharose
-
sucrose density gradient, green A-agarose, Mono Q, Superose-6
-
sucrose gradient
-
the recombinant protein is purified to homogeneity by using ion-exchange and size-exclusion chromatography
thermosomes are purified using a Resource-Q anion exchange column
using cation exchange chromatography and gel filtration
-
using two ion-exchange dimensions followed by gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning of GroEL wild-type and mutants in Escherichia coli strain DH5alpha, recombinant expression in Escherichia coli strain BL21(DE3), co-expression with cofactor GroES
-
expressed in Escherichia coli
-
expressed in Escherichia coli, alpha and beta subunits
-
expression Escherichia coli
expression in Escherichia coli
expression in Escherichia coli BL21
expression in Escherichia coli; expression in Escherichia coli
full-length u45 mutant and wild-type hsp90a are amplified by RT-PCR, cloned into the the TOPO vector and subcloned by PCR into pCS2myc+
-
full-length wild-type E. coli GroEL is expressed in Escherichia coli strain W3110 bearing a multicopy of the noninducible pOF39 plasmid
-
gene groEL, DNA and amino acid sequence determination and analysis, recombinant expression of the His-tagged enzyme in Escherichia coli strain BL21(DE3)
GroEL in pTRC99a is overexpressed in Escherichia coli DH5alpha cells; the N-cpn20 and C-cpn20 homologs of mature wild type cpn20 are cloned individually into the pGEM T-easy vector, the constructs are further cloned into pET22b+
-
GroEL mutants are constructed in a pCH vector backbone, chaperonin constructs for in vivo co-expression are prepared by inserting the fragments into the pOFXtac-SL2 vector, furthermore the vectors pCH-L16-GFP and pBAD18 are used
-
into the pET24d vector for expression in Escherichia coli BL21DE3 cells
-
into the pET3a vector for expression in Escherichia coli BL21DE3 cells, the pOFX HSP60wt-HSP10wt plasmid is used for establishing a bacterial in vivo system
-
recombinant expression of chaperonin GroEL in Escherichia coli cells bearing the multicopy plasmid pGroESL
-
the amplified structural gene is expressed in Escherichia coli BLR DE3 using vector pET23a+
the chaperonin subunit is expressed in Escherichia coli strain BL21DE3 cells using the expression plasmid pK1Ebeta; the chaperonin subunits are expressed in Escherichia coli strain BL21DE3 cells using the expression plasmid pK1Ealpha and pK1Ealpha2 for the alpha-subunit mutant and the wild-type alpha-subunit
-
the Pyrococcus furiosus chaperonin gene is amplified by PCR from genomic DNA, cloned into pET21, and expressed in Escherichia coli BL21-Codonplus DE3-RIL cells
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
GroEL/GroES overexpression doubles the number of accumulating mutations, and promotes the folding of enzyme variants carrying mutations in the protein core and/or mutations with higher destabilizing effects
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
R285H
-
no significant difference in the refolding activity between mutant and wild-type GroEL
T93I
-
no significant difference in the refolding activity between mutant and wild-type GroEL. No significant difference in inorganic phosphate release between mutant and wild-type enzyme at 10C, 37C and 50C
slou45
-
the ATPase activity of Hsp90 is abrogated by the slou45 mutation
A399T
-
site-diretected mutagenesis, the mutation weakens the affinity for GroES by about 90fold
A92T
-
site-diretected mutagenesis, the mutation weakens the affinity for GroES by about 1600fold
C138S/C458S/C519S/D83C/K327C
conformational change, in reduced state similar ATP hydrolysis and ATP binding to wild-type GroEL
D115N
-
site-diretected mutagenesis, the mutation weakens the affinity for GroES by about 50fold
D398K
-
block of ATP hydrolysis
D52A
-
site-directed mutagenesis, ATPase activity of the mutant is 80% reduced compared to the wild-type GroEL
D52A/D398A
E191G
-
site-diretected mutagenesis, the mutation weakens the affinity for GroES by about 300fold
E461K
-
site-directed mutagenesis, the inactive mutant of GroEL has a rearranged inter-ring interface, the normal 1:2 contacts of apposed equatorial domains in wild-type GroEL are replaced by 1:1 contacts in the mutant in the interfaces