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Information on EC 2.8.3.12 - glutaconate CoA-transferase and Organism(s) Acidaminococcus fermentans and UniProt Accession Q59111
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2.8.3.12 glutaconate CoA-transferaseIUBMB Comments
Glutarate, (R)-2-hydroxyglutarate, propenoate and propanoate, but not (Z)-glutaconate, can also act as acceptors.
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Word Map
- 2.8.3.12
- fermentans
-
acidaminococcus
-
r-2-hydroxyglutarate
-
r-2-hydroxyglutaryl-coa
- dehydration
- crotonyl-coa
- coash
- dehydratase
- thiolester
- hydroxyglutarate
- glutaconyl-coa
-
coa-ester
- aminobutyricum
-
anhydride
- 4-hydroxybutyrate
- glutaryl-coa
- propionicum
- beta-subunit
The taxonomic range for the selected organisms is: Acidaminococcus fermentans
The expected taxonomic range for this enzyme is: Bacteria, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Synonyms
glutaconate coa-transferase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
(E)-glutaconate CoA-transferase
-
-
-
-
coenzyme A-transferase, glutaconate
-
-
-
-
GCT
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
acetyl-CoA + (E)-glutaconate = acetate + glutaconyl-1-CoA
ping-pong mechanism
-
acetyl-CoA + (E)-glutaconate = acetate + glutaconyl-1-CoA
reaction mechanism
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
coenzyme A transfer
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -
SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:(E)-glutaconate CoA-transferase
Glutarate, (R)-2-hydroxyglutarate, propenoate and propanoate, but not (Z)-glutaconate, can also act as acceptors.
CAS REGISTRY NUMBER
COMMENTARY
79078-99-2
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + acrylate
acrylyl-CoA + acetate
-
poor substrate
-
-
?
acetyl-CoA + crotonate
crotonyl-CoA + acetate
-
poor substrate
-
-
?
acetyl-CoA + isocrotonate
isocrotonyl-CoA + acetate
-
poor substrate
-
-
?
glutaconyl-CoA + (S)-2-hydroxyglutarate
glutaconate + (S)-2-hydroxyglutaryl-CoA
-
at 60% of the rate with (R)-isomer or glutarate
-
-
?
glutaconyl-CoA + 3-hydroxyglutarate
glutaconate + 3-hydroxyglutaryl-CoA
-
at 20% of the rate with glutarate
-
-
?
glutaconyl-CoA + adipate
glutaconate + adipyl-CoA
-
at 60% of the rate with glutarate
-
-
?
glutaconyl-CoA + glutarate
glutaconate + glutaryl-CoA
-
very good substrate
-
?
glutaconyl-CoA + propionate
propanoyl-CoA + glutaconate
-
moderate substrate
-
-
?
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
-
-
-
?
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
-
-
-
?
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
-
-
-
r
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
-
-
-
?
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
-
-
only 1-isomer is formed
?
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
-
very good substrate, broad substrate specificity
-
?, r
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
-
removal of 3'-phospho group from acetyl-CoA leads to reduced activity
-
?, r
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
-
involved in glutamate fermentation via hydroxyglutarate pathway, activates glutaconate to glutaconyl-CoA prior to glutaconate decarboxylation
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
-
-
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
-
-
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
-
-
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
-
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
-
-
in vitro formation of both isomers: (R)-2-hydroxyglutaryl-1-CoA and (R)-2-hydroxyglutaryl-5-CoA, the 1-isomer is kinetically favoured, the 5-isomer is thermodynamically favoured
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
-
-
in vitro formation of both isomers: (R)-2-hydroxyglutaryl-1-CoA and (R)-2-hydroxyglutaryl-5-CoA, the 1-isomer is kinetically favoured, the 5-isomer is thermodynamically favoured, product in vivo, 5-isomer only occurs in vitro, (R)-2-hydroxyglutaryl-1-CoA is an excellent substrate in the reverse reaction
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
-
in the course of glutamate fermentation
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
-
in the course of glutamate fermentation
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
-
in the course of glutamate fermentation
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
involved in the conversion of (R)-2-hydroxyglutarate to crotonyl-CoA in the pathway of glutamate fermentation
-
?
glutaconyl-CoA + (R)-2-hydroxyglutarate
glutaconate + (R)-2-hydroxyglutaryl-CoA
-
-
formation of the 5-isomer, not of the 1-isomer
?
glutaconyl-CoA + (R)-2-hydroxyglutarate
glutaconate + (R)-2-hydroxyglutaryl-CoA
-
very good substrate
generation of both isomers: (R)-2-hydroxyglutaryl-1-CoA and (R)-2-hydroxyglutaryl-5-CoA
?
glutaryl-CoA + acetate
glutarate + acetyl-CoA
-
-
?
glutaryl-CoA + acetate
glutarate + acetyl-CoA
-
exchange of oxygen atoms between the substrates and the catalytic residue betaE-54, exchange is site-specific, reversible and requires both substrates, catalytic mechanism
-
r
additional information
?
-
-
wild-type enzyme has significant hydrolase activity using acetyl-CoA as substrate, but no hydrolysis of glutaryl-CoA
-
-
?
additional information
?
-
-
residue E-54 of the subunit GctB is directly involved in catalysis by formation of a CoASH ester intermediate
-
-
?
additional information
?
-
residue E-54 of the small subunit GctB is directly involved in catalysis
-
-
?
additional information
?
-
-
residue E-54 of the small subunit GctB is directly involved in catalysis
-
-
?
additional information
?
-
-
no substrates: (Z)-glutaconate, acetyl-4'-phosphopantetheine, C4-dicarboxylic acids, reverse reaction not with glutamate, 2-oxoglutarate, succinate, malate or citrate
-
-
?
additional information
?
-
-
GCT structure, the active site is located at the interface of subunits A and B and is formed by loops of both subunits
-
-
?
additional information
?
-
-
catalytic residue is E-54 of subunit B, catalytic mechanism
-
-
?
additional information
?
-
-
catalytic residue is E-54 of subunit B, catalytic mechanism
-
-
?
additional information
?
-
-
catalytic residue is E-54 of subunit B, catalytic mechanism
-
-
?
additional information
?
-
-
not involved in the dehydration of (R)-2-hydroxyglutarate to glutaconate
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
-
involved in glutamate fermentation via hydroxyglutarate pathway, activates glutaconate to glutaconyl-CoA prior to glutaconate decarboxylation
-
?
additional information
?
-
-
not involved in the dehydration of (R)-2-hydroxyglutarate to glutaconate
-
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
-
in the course of glutamate fermentation
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
-
in the course of glutamate fermentation
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
-
in the course of glutamate fermentation
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
involved in the conversion of (R)-2-hydroxyglutarate to crotonyl-CoA in the pathway of glutamate fermentation
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4-hydroxymercuribenzoate
-
0.2 mM, at 37°C, 2 min, 80% loss of activity, 2-mercaptoethanol partially restores activity
additional information
-
not inhibited by 2 mM iodoacetate, 2 mM iodoacetamide or 1 mM 5,5-dithiobis(2-nitrobenzoate)
-
NaBH4
-
10 mM, in presence of 0.4 mM glutyral-CoA, at 25°C, 30 min, almost complete inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.035
(R)-2-hydroxyglutaryl-1-CoA
-
pH 7, 37°C, cosubstrate acetate
0.064
glutaryl-CoA
-
pH 7, cosubstrate acetate, 65 kDa GctF fusion protein
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
39
glutaryl-CoA
-
pH 7, cosubstrate acetate, 65 kDa GctF fusion protein
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
37
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
5-part of the hydroxyglutarate operon with gctAB genes encoding Gct, ATCC 25085
UniProt
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
29000
29018
-
alpha4beta4, 4 * 35573 + 4 * 29018, MALDI-TOF mass spectrometry
29168
alpha4beta4, 4 * 35725 + 4 * 29168, GctA and GctB, calculated from the DNA sequences
35000
-
alpha4beta4, 4 * 35000 + 4 * 29000, structure and association of the subunits A and B
35573
-
alpha4beta4, 4 * 35573 + 4 * 29018, MALDI-TOF mass spectrometry
35725
alpha4beta4, 4 * 35725 + 4 * 29168, GctA and GctB, calculated from the DNA sequences
36000
29000
-
alpha4beta4, 4 * 36000 + 4 * 29000, GctA and GctB, SDS-PAGE
29000
alpha4beta4, 4 * 36000 + 4 * 29000, GctA and GctB, SDS-PAGE
29000
-
alpha4beta4, 4 * 35000 + 4 * 29000, structure and association of the subunits A and B
36000
-
alpha4beta4, 4 * 36000 + 4 * 29000, GctA and GctB, SDS-PAGE
36000
alpha4beta4, 4 * 36000 + 4 * 29000, GctA and GctB, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
octamer
octamer
-
alpha4beta4, 4 * 36000 + 4 * 29000, GctA and GctB, SDS-PAGE
octamer
alpha4beta4, 4 * 36000 + 4 * 29000, GctA and GctB, SDS-PAGE
octamer
alpha4beta4, 4 * 35725 + 4 * 29168, GctA and GctB, calculated from the DNA sequences
octamer
-
alpha4beta4, 4 * 35573 + 4 * 29018, MALDI-TOF mass spectrometry
octamer
-
alpha4beta4, 4 * 35000 + 4 * 29000, structure and association of the subunits A and B
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
native and recombinant enzyme, expressed in Escherichia coli
recombinant GCT, expressed in Escherichia coli, sitting-drop vapour diffusion method
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
E54A
E54D
E54N
E54Q
E64A
-
mutation in the subunit GctB, 30% as active as wild-type enzyme
E54A
-
mutation in the subunit GctB, inactive mutant
E54A
-
mutant without CoA-transferase or acyl-CoA hydrolase activity
E54D
-
replacement of the catalytic glutamate by aspartate converts the mutant enzyme to a thiol ester hydrolase
E54D
-
mutation decreases CoA-transferase and increases acyl-CoA hydrolase activity, mechanism, mutant catalyses the hydrolysis of glutaryl-CoA, acetyl-CoA and 3-butenoyl-CoA, mutant Escherichia coli strain shows reduced growth rate after induction
E54D
-
above 7% as active as wild-type enzyme, mutant with changed catalytic mechanism without CoASH intermediate, the Escherichia coli strain producing mutant enzyme shows a lower growth rate and reduced amount of recombinant enzyme
E54N
-
mutant without CoA-transferase or acyl-CoA hydrolase activity
E54Q
-
1% as active as wild-type enzyme, by incubating with both substrates for 20 h at room temperature, glutamine is completely converted to glutamate yielding a fully active CoA-transferase
E54Q
-
mutant activity increases from 1% to almost 100% upon incubation with acetyl-CoA and glutaconate at 37°C within 40 h, the substrates induce the conversion of the mutant glutamine residue into the glutamate residue of the wild-type enzyme
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, crystalline suspension in ammonium sulfate, 2 years, stable
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
betaE54D mutant enzyme and 65 kDa GctF fusion protein, expressed in Escherichia coli
-
native and recombinant enzyme, expressed in Escherichia coli
recombinant wild-type and betaE54D mutant enzyme, expressed in Escherichia coli
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gctAB genes encoding the two subunits GctA and GctB are cloned, sequenced and overexpressed together in Escherichia coli DH5alpha, the two genes are located at the beginning of the hydroxyglutarate operon, GctA: 320 amino acids protein, GctB: 266 amino acids protein
overexpression of betaE54N and betaE54A mutant enzyme in Escherichia coli, fusion of the genes gctA and gctB, encoding the 2 subunits of Gct, yields GctF, which is expressed in Escherichia coli as a 65 kDa protein with 30% of wild-type activity
-
overexpression of wild-type and betaE54D mutant enzyme in Escherichia coli
-
wild-type and mutant genes gctA and gctB encoding the two subunits GctA and GctB are cloned and expressed together in Escherichia coli XL1-Blue
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Mack, M.; Buckel, W
Conversion of glutaconate CoA-transferase from Acidaminococcus fermentans into an acyl-CoA hydrolase by site-directed mutagenesis
FEBS Lett.
405
209-212
1997
Acidaminococcus fermentans
Buckel, W.; Dorn, U.; Semmler, R.
Glutaconate CoA-transferase from Acidaminococcus fermentans
Eur. J. Biochem.
118
315-321
1981
Acidaminococcus fermentans, [Clostridium] symbiosum, [Clostridium] sporosphaeroides, no activity in Clostridium tetanomorphum, [Clostridium] symbiosum HB 25, no activity in Clostridium tetanomorphum H1
Klees, A.G.; Buckel, W.
Synthesis and properties of (R)-2-hydroxyglutaryl-1-CoA. (R)-2-hydroxyglutaryl-5-CoA, an erroneous product of glutaconate CoA-transferase
Biol. Chem. Hoppe-Seyler
372
319-324
1991
Acidaminococcus fermentans
Mack, M.; Bendrat, K.; Zelder, O.; Eckel, E.; Linder, D.; Buckel, W.
Location of the two genes encoding glutaconate coenzyme A-transferase at the beginning of the hydroxyglutarate operon in Acidaminococcus fermentans
Eur. J. Biochem.
226
41-51
1994
Acidaminococcus fermentans (Q59111 and Q59112), Acidaminococcus fermentans
Mack, M.; Buckel, W.
Identification of glutamate beta54 as the covalent-catalytic residue in the active site of glutaconate CoA-transferase from Acidaminococcus fermentans
FEBS Lett.
357
145-148
1995
Acidaminococcus fermentans
Jacob, U.; Mack, M.; Clausen, T.; Huber, R.; Buckel, W.; Messerschmidt, A.
Glutaconate CoA-transferase from Acidaminococcus fermentans: the crystal structure reveals homology with other CoA-transferases
Structure
5
415-426
1997
Acidaminococcus fermentans
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