Information on EC 6.4.1.4 - methylcrotonoyl-CoA carboxylase

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

EC NUMBER
COMMENTARY hide
6.4.1.4
-
RECOMMENDED NAME
GeneOntology No.
methylcrotonoyl-CoA carboxylase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + 3-methylcrotonoyl-CoA + HCO3- = ADP + phosphate + 3-methylglutaconyl-CoA
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
carboxylation
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
L-leucine degradation I
-
-
Metabolic pathways
-
-
Valine, leucine and isoleucine degradation
-
-
leucine metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
3-methylcrotonoyl-CoA:carbon-dioxide ligase (ADP-forming)
A biotinyl-protein.
CAS REGISTRY NUMBER
COMMENTARY hide
9023-95-4
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
IVS
-
-
Manually annotated by BRENDA team
Columbia ecotype
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
carrot
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
pregnant mice
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain PAo1 Smr ins-atuF
-
-
Manually annotated by BRENDA team
Wistar strain
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3-methylcrotonyl-CoA + ATP + HCO3-
3-methylglutaconyl-CoA + ADP + phosphate
show the reaction diagram
ATP + (2Z)-3-ethylcrotonoyl-CoA
ADP + phosphate + 3-ethylglutaconyl-CoA
show the reaction diagram
-
12% of the activity relative to 3-methylcrotonoyl-CoA
-
-
-
ATP + 3-methylcrotonoyl-CoA + HCO3-
?
show the reaction diagram
ATP + 3-methylcrotonoyl-CoA + HCO3-
ADP + phosphate + 3-methylglutaconyl-CoA
show the reaction diagram
ATP + 3-methylcrotonyl-CoA + HCO3-
ADP + 3-methylglutaconyl-CoA + phosphate
show the reaction diagram
ATP + 3-methylcrotonyl-CoA + HCO3-
ADP + phosphate + 3-methylglutaconyl-CoA
show the reaction diagram
ATP + acetoacetyl-CoA + HCO3-
?
show the reaction diagram
ATP + acetyl-CoA + HCO3-
ADP + malonyl-CoA + phosphate
show the reaction diagram
-
-
-
-
r
ATP + crotonoyl-CoA + HCO3-
?
show the reaction diagram
ATP + crotonyl-CoA + HCO3-
ADP + phosphate + glutaconyl-CoA
show the reaction diagram
glutaconyl-CoA + H+
crotonyl-CoA + CO2
show the reaction diagram
-
-
-
r
UTP + 3-methylcrotonoyl-CoA + HCO3-
UDP + phosphate + 3-methylglutaconyl-CoA
show the reaction diagram
-
at 62% of the activity relative to ATP
-
-
-
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
3-methylcrotonyl-CoA + ATP + HCO3-
3-methylglutaconyl-CoA + ADP + phosphate
show the reaction diagram
-
mitochondrial leucine catabolic pathway
-
?
ATP + 3-methylcrotonoyl-CoA + HCO3-
?
show the reaction diagram
ATP + 3-methylcrotonoyl-CoA + HCO3-
ADP + phosphate + 3-methylglutaconyl-CoA
show the reaction diagram
ATP + 3-methylcrotonyl-CoA + HCO3-
ADP + 3-methylglutaconyl-CoA + phosphate
show the reaction diagram
ATP + 3-methylcrotonyl-CoA + HCO3-
ADP + phosphate + 3-methylglutaconyl-CoA
show the reaction diagram
ATP + acetyl-CoA + HCO3-
ADP + malonyl-CoA + phosphate
show the reaction diagram
-
-
-
-
r
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
biotin
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Li+
-
relative activation by monovalent cations in decreasing order: K+/ NH4+, Na+, Li+
Na+
-
relative activation by monovalent cations in decreasing order: K+/ NH4+, Na+, Li+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2E)-3-Ethylcrotonoyl-CoA
-
competitive with respect to 3-methylcrotonoyl-CoA
(2Z)-geranoyl-CoA
-
competitive with respect to 3-methylcrotonoyl-CoA
(E)-2-methyl-2-butenoyl-CoA
-
weak
acetoacetyl-CoA
ADP
-
ADP and phosphate inhibit enzyme activity in an additive mode
ATP
-
-
Avidin
-
irreversible inhibition
-
Butanoyl-CoA
iodoacetamide
-
3-methylcrotonoyl-CoA protects against inactivation
isobutanoyl-CoA
-
weak
n-Hexanoyl-CoA
-
weak
p-hydroxymercuribenzoate
palmitoyl-CoA
-
25% inhibition
pentanoyl-CoA
-
weak
Phenylglyoxal
phosphate
-
ADP and phosphate inhibit enzyme activity in an additive mode
propanoyl-CoA
-
competitive with respect to 3-methylcrotonoyl-CoA
Streptavidin
-
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
biotin
additional information
-
activity is stimulated by continuous darkness
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.022
(2Z)-3-ethylcrotonoyl-CoA
-
-
0.138 - 0.225
(E)-crotonoyl-CoA
0.0098 - 31
3-methylcrotonoyl-CoA
0.074
3-methylcrotonyl-CoA
-
37C, pH 8.0, recombinant enzyme
0.01 - 21
ATP
2.8
bicarbonate
-
experimental conditions not available
0.11
crotonoyl-CoA
-
-
0.0008 - 2.2
HCO3-
additional information
additional information
-
MCCase activity shows sigmoidal kinetics for all the substrates and non-Michaelis-Menten kinetics, overview
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.43 - 2.76
3-methylcrotonoyl-CoA
4
3-methylcrotonyl-CoA
Homo sapiens
-
37C, pH 8.0, recombinant enzyme
4
ATP
Homo sapiens
-
37C, pH 8.0, recombinant enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.39 - 5.36
3-methylcrotonoyl-CoA
2462
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.1182
-
-
0.206
-
-
0.2062
-
-
0.3367
-
-
3.7
-
-
3.84
-
kidney enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5 - 9
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7.5: about 30% of maximal activity, 9.0: about 85% of maximal activity
7.9 - 8.3
-
-
8
-
in presence of Mg2+ and K+
8 - 8.5
8 - 8.3
-
-
8 - 8.5
8 - 8.2
-
-
8 - 8.5
8
-
-
8.5
-
native and recombinant enzyme
9
-
recombinant enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8 - 10.1
-
6.8: more than 70% of maximal activity, 9.5: more than 70% of maximal activity, 10.1: 33% of maximal activity
8 - 9
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pH 6.0: no activity, 8.0: maximal activity, pH 9.0: 50% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
recombinant enzyme
37
-
native and recombinant enzyme
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23 - 50
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23C: 60% of maximal activity, 30C: 75% of maximal activity, 50C: 15% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
presence of mRNA as well as protein of both subunits of 3-methylcrotonyl-CoA carboxylase in ependymocyte, microglial cell and oligodendrocyte cultures derived from the brains of newborn rats
Manually annotated by BRENDA team
presence of mRNA as well as protein of both subunits of 3-methylcrotonyl-CoA carboxylase in ependymocyte, microglial cell and oligodendrocyte cultures derived from the brains of newborn rats
Manually annotated by BRENDA team
-
only trace levels
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
presence of mRNA as well as protein of both subunits of 3-methylcrotonyl-CoA carboxylase in ependymocyte, microglial and oligodendrocyte cultures derived from the brains of newborn rats
Manually annotated by BRENDA team
presence of mRNA as well as protein of both subunits of 3-methylcrotonyl-CoA carboxylase in ependymocyte, microglial cell and oligodendroglial cell cultures derived from the brains of newborn rats
Manually annotated by BRENDA team
-
only trace levels
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
11624
-
1 * 65000 + 1 * 36000 + 1 * 25000 + 1 * 14000, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, SDS-PAGE, 1 * 64300 + 1 * 38902 + 1 * 14133 + 1 * 11624, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, amino acid sequence
14000
-
1 * 65000 + 1 * 36000 + 1 * 25000 + 1 * 14000, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, SDS-PAGE, 1 * 64300 + 1 * 38902 + 1 * 14133 + 1 * 11624, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, amino acid sequence
14133
-
1 * 65000 + 1 * 36000 + 1 * 25000 + 1 * 14000, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, SDS-PAGE, 1 * 64300 + 1 * 38902 + 1 * 14133 + 1 * 11624, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, amino acid sequence
25000
-
1 * 65000 + 1 * 36000 + 1 * 25000 + 1 * 14000, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, SDS-PAGE, 1 * 64300 + 1 * 38902 + 1 * 14133 + 1 * 11624, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, amino acid sequence
36000
-
1 * 65000 + 1 * 36000 + 1 * 25000 + 1 * 14000, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, SDS-PAGE, 1 * 64300 + 1 * 38902 + 1 * 14133 + 1 * 11624, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, amino acid sequence
38902
-
1 * 65000 + 1 * 36000 + 1 * 25000 + 1 * 14000, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, SDS-PAGE, 1 * 64300 + 1 * 38902 + 1 * 14133 + 1 * 11624, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, amino acid sequence
53000
-
4 * 74000, biotin-containing subunit, + 4 * 53000, biotin-free subunit, SDS-PAGE
54000
-
4 * 76000, biotin-containing subunit, + 4 * 54000, biotin-free subunit, SDS-PAGE
58500
-
6 * 58500 + 6 * 80000, SDS-PAGE
61000
-
x * 61000, biotin-free subunit A, + x * 73500, biotin-containing subunit B
62000
-
x * 62000, biotin-free subunit, + x * 80000, biotin-containing subunit, SDS-PAGE
64300
-
1 * 65000 + 1 * 36000 + 1 * 25000 + 1 * 14000, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, SDS-PAGE, 1 * 64300 + 1 * 38902 + 1 * 14133 + 1 * 11624, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, amino acid sequence
65000
-
1 * 65000 + 1 * 36000 + 1 * 25000 + 1 * 14000, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, SDS-PAGE, 1 * 64300 + 1 * 38902 + 1 * 14133 + 1 * 11624, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, amino acid sequence
72000
-
1 * 60000 + 1 * 72000, SDS-PAGE, Western Blot
73500
-
x * 61000, biotin-free subunit A, + x * 73500, biotin-containing subunit B
490000
-
gel filtration
500000
520000 - 580000
-
gel filtration
530000
-
gel filtration
660000
-
-
700000 - 760000
-
-
760000
-
analytical ultracentrifugation
800000 - 900000
800000
835000
900000
nondenaturing PAGE, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dodecamer
heterodimer
heteromer
heterotetramer
-
co-expressed subunits form an (alpha-beta)4 complex that suffer a modification of its oligomerization
homodimer
-
alphabeta
multimer
-
x * 60000 + 6 * 80000, 3-methylcrotonyl-CoA carboxylase is a heteromultimeric complex (MW: 500000 Da-800000 Da) that is composed of alpha and beta subunits (ratio 1:1) which are encoded by distinct genes. MCCCalpha is the larger subunit that has 725 amino acids, which contains the Met-Lys-Met biotin attachment domain near its COOH end. Human MCCCbeta is the small subunit that is composed of 563 amino acids, which functions as a carboxyltransferase, SDS-PAGE
octamer
polymer
x * 85000 + x * 60000, heteromeric enzyme composed of biotin-containing MCC-A and non-biotin-containing MCC-B subunits
tetramer
-
1 * 65000 + 1 * 36000 + 1 * 25000 + 1 * 14000, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, SDS-PAGE, 1 * 64300 + 1 * 38902 + 1 * 14133 + 1 * 11624, alpha GcdA, beta GcdB, gamma GcdC, delta GcdD, amino acid sequence
additional information
-
heteromeric enzyme composed of 2 kinds of subunits, biotinylated MCC-A and non-, biotinylated MCC-B
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structures diffracted to 1.5 A resolution of the Pseudomonas aeruginosa MCC (PaMCC) holoenzyme are shown, alone and in complex with coenzyme A. The structures show that the architecture and overall shape of PaMCC are strikingly different when compared to propionyl-CoA carboxylase; crystal structures diffracted to 1.5 A resolution of the Pseudomonas aeruginosa MCC (PaMCC) holoenzyme are shown, alone and in complex with coenzyme A. The structures show that the architecture and overall shape of PaMCC are strikingly different when compared to propionyl-CoA carboxylase
crystal structures diffracted to 1.5 A resolution of the Pseudomonas aeruginosa MCC (PaMCC) holoenzyme are shown, alone and in complex with coenzyme A. The structures show that the architecture and overall shape of PaMCC are strikingly different when compared to propionyl-CoA carboxylase
-
crystal structures of the Pseudomonas aeruginosa MCC (PaMCC) holoenzyme, alone and in complex with coenzyme A are shown at 2.9 and 3.5 A resolution respectively. The structures show that the architecture and overall shape of PaMCC are strikingly different when compared to propionyl-CoA carboxylase
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
-
1 min, 50% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
MCCase extracted with 0.1 M Tris-HCl pH 8.0 is relatively unstable, enzyme is completely inactive 2-3 days after extraction, changing of pH or addition of bovine serum albumin fails to stabilize the enzyme
-
very unstable, even in presence of potential stabilizing agents such as dithiothreitol, EDTA and glycerol 50% of beta-MCC activity is lost in 24 h
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 4 months, less than 20% loss of activity
-
-20C, for at least 3 months without loss of activity
-
-20C, purified rnzyme can be stored up to 6 months with minimal loss of activity
-
-80C, approximately 25% loss of activity per month
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
kidney enzyme
-
recombinant alpha-subunit
-
recombinant His-tagged MCCase from Escherichia coli strain BL21 by avidin affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cDNA cloned and expressed in Escherichia coli
-
decarboxylase operon cloned and sequenced, overproduced in Escherichia coli
-
expressed in Escherichia coli
-
expression as fusion proteins in human fibroblasts
-
expression of His-tagged MCCase in Escherichia coli strain BL21
-
MCCase subunits are co-expressed in Escherichia coli in the same plasmid and purified in one step by affinity chromatography using the LiuD-His tag protein interacting with the LiuB-S tag recombinant protein
-
reverse transcriptase PCR
-
singlecopy gene encoding MCC-A subunit isolated nd characterized
-
the alpha and beta subunits of PaMCC are co-expressed in Escherichia coli as His-tagged fusion proteins
-
use of baculovirus system to express functionally active human recombinant 3-methylcrotonyl-CoA carboxylase
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
M169K
-
mutant with lack of carboxylation activity
R338Q
-
mutant with lack of carboxylation activity
R338S
-
mutant with lack of carboxylation activity
R385S
-
human mutant leading to methylcrotonylglycinuria
G46E
-
the mutation of the MCCA gene is associated with methylcrotonylglycinuria
G517R
-
the mutation of the MCCB gene is associated with methylcrotonylglycinuria
L355F
-
the mutation of the MCCB gene is associated with methylcrotonylglycinuria
Q477R
-
the mutation of the MCCB gene is associated with methylcrotonylglycinuria
R385S
-
mutant leading to biochemical abnormalities and clinical symptoms in heterozygous individuals
Y520S
-
the mutation of the MCCB gene is associated with methylcrotonylglycinuria
F417Y
-
beta-subunit mutant retains activity, kcat/Km increased compared to wild-type, kcat (3-methylcrotonoyl-CoA) increased compared to wild-type, Km (3-methylcrotonoyl-CoA) increased compared to wild-type
F417Y/G423A
-
beta-subunit mutant retains activity, kcat/Km similar to wild-type, kcat (3-methylcrotonoyl-CoA) increased compared to wild-type, Km (3-methylcrotonoyl-CoA) increased compared to wild-type
G423A
-
beta-subunit mutant retains activity, kcat/Km decreased compared to wild-type, kcat (3-methylcrotonoyl-CoA) decreased compared to wild-type, Km (3-methylcrotonoyl-CoA) slightly decreased compared to wild-type
R51A
-
beta-subunit mutant retains activity, kcat/Km decreased compared to wild-type, kcat (3-methylcrotonoyl-CoA) decreased compared to wild-type, Km (3-methylcrotonoyl-CoA) slightly increased compared to wild-type
S187A
-
beta-subunit mutant shows no enzymatic activity
Y422D
-
beta-subunit mutant shows no enzymatic activity
additional information
-
the IVS7-1GNA splice site mutation of the MCCB gene is associated with methylcrotonylglycinuria and shows 31fold reduced activity compared to the wild type
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
MCCase is of significance in comprehending how the mevalonate shunt can divert carbon away from the biosynthesis of isoprenoids, such as cholesterol, which has major implications in the prevention of vascular degenerate diseases
medicine
Show AA Sequence (1213 entries)
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