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Information on EC 5.4.99.2 - methylmalonyl-CoA mutase and Organism(s) Homo sapiens and UniProt Accession P22033

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EC Tree
     5 Isomerases
         5.4 Intramolecular transferases
             5.4.99 Transferring other groups
                5.4.99.2 methylmalonyl-CoA mutase
IUBMB Comments
Requires a cobamide coenzyme.
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This record set is specific for:
Homo sapiens
UNIPROT: P22033
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Word Map
The taxonomic range for the selected organisms is: Homo sapiens
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Synonyms
methylmalonyl-coa mutase, methylmalonyl coa mutase, l-methylmalonyl-coa mutase, methylmalonyl coenzyme a mutase, sleeping beauty mutase, mitochondrial methylmalonyl-coa mutase, mmcm-1, (s)-methylmalonyl-coa mutase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Methylmalonyl coenzyme A mutase
-
mitochondrial methylmalonyl-CoA mutase
-
(R)-2-methyl-3-oxopropanoyl-CoA CoA-carbonylmutase
-
-
-
-
(S)-Methylmalonyl-CoA mutase
-
-
-
-
MCB-beta
-
-
-
-
MCM-alpha
-
-
-
-
MCM-beta
-
-
-
-
Methylmalonyl CoA mutase
-
-
-
-
Methylmalonyl coenzyme A carbonylmutase
-
-
-
-
Methylmalonyl coenzyme A mutase
-
-
-
-
Methylmalonyl-CoA-carbonyl mutase
-
-
-
-
Mutase, methylmalonyl coenzyme A
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(R)-methylmalonyl-CoA = succinyl-CoA
show the reaction diagram
mechanism, overview, a gradual weakening of the electrostatic energy between the protein and the ribose leads to a progressive increase in the activation energy barrier for Co-C bond homolysis, key role for the conserved polar glutamate residue in controlling the initial generation of radical species
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
isomerization
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
(R)-methylmalonyl-CoA CoA-carbonylmutase
Requires a cobamide coenzyme.
CAS REGISTRY NUMBER
COMMENTARY hide
9023-90-9
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-2-methylmalonyl-CoA
succinyl-CoA
show the reaction diagram
-
-
-
?
(R)-methylmalonyl-CoA
succinyl-CoA
show the reaction diagram
(R)-2-Methyl-3-oxopropanoyl-CoA
?
show the reaction diagram
-
-
-
-
?
(R)-2-methylmalonyl-CoA
succinyl-CoA
show the reaction diagram
(R/S)-2-methyl-3-oxopropanoyl-CoA
succinyl-CoA
show the reaction diagram
-
-
-
-
?
(S)-methylmalonyl-CoA
succinyl-CoA
show the reaction diagram
-
-
-
-
?
ethylmalonyl-CoA
methylsuccinyl-CoA
show the reaction diagram
-
reacts about 1000-10000 times moreslowly than the natural substrate
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(R)-methylmalonyl-CoA
succinyl-CoA
show the reaction diagram
(R)-2-Methyl-3-oxopropanoyl-CoA
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
adenosylcobalamin
Cobalamin
essential to the function of methylmalonyl-CoA mutase, cobalamin is conversed to adenosylcobalamin
adenosylcobalamin
-
MCM activity with adenosylcobalamin increases linearly during the first 3-4 min
Cobalamin
-
dependent on, 2 mol of cobalamin bound per mol of enzyme, are covalently attached
cobamide
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
required
Cl-
-
stimulates
PO43-
-
required, maximal activity at 50 mM
SO42-
-
stimulates
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(R)-2-methyl-3-oxopropanoyl-CoA
-
substrate inhibition
Ca2+
-
at high concentrations
Cyclopropylcarbonyl-CoA carboxylate
-
reversible mixed-type inhibition
methylenecyclopropylacetyl-CoA
-
reversible mixed-type inhibition
methylmalonyl-CoA
-
reversible mixed-type inhibition
Mg2+
-
at high concentrations
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
hMMAA
hMCM loses activity during catalysis, but the interaction with human MMAA (hMMAA), a GTPase protein, avoids this loss or restores hMCM activity. Formation and accumulation of OH2Cbl, the oxidized form of cofactor AdoCbl formed during catalysis, is the cause of hMCM inactivation. The complex formation of hMCM/hMMAA decreases the rate of oxidized cofactor formation, protecting the hMCM enzyme. hMMAA is able to remove the damaged cofactor through GTP hydrolysis.a modification in the kinetic parameters of hMCM in presence of hMMAA is observed. No activity is recovered in the presence of hMMAAwith GMPPNP, confirming the need of GTP hydrolysis for this cofactor removal. In vivo localization of hMMAA and its colocalization with hMCM in human fibroblasts mitochondria. hMMAAis localized in both the cytoplasm and mitochondria but in less abundance and distribution than enzyme hMCM
-
adenosylcobalamin
-
required
methylmalonic acidemia protein
-
after 60 min of reaction, when MCM is inactive, the addition of methylmalonic acidemia protein increases the enzymatic activity through GTP hydrolysis, indicating reactivation of MCM by exchange of the damaged cofactor. Methylmalonic acidemia protein acts as a chaperone of human MCM
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.5
(R)-2-methyl-3-oxopropanoyl-CoA
-
-
0.06 - 0.2
(R/S)-2-methyl-3-oxopropanoyl-CoA
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.012 - 3.95
(R)-methylmalonyl-CoA
150 - 283
(R/S)-2-methyl-3-oxopropanoyl-CoA
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.33
-
-
67.4
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.5
assay at
6.5 - 8
-
-
7 - 9
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
assay at
25
assay at
37
assay at
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
peripheral lymphocytes
Manually annotated by BRENDA team
-
peripheral blood lymphocyte
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
enzyme hMCM preferably locates in mitochondria, with small amounts of protein in the cytoplasm
Manually annotated by BRENDA team
additional information
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
MUTA_HUMAN
750
0
83134
Swiss-Prot
Mitochondrion (Reliability: 3)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
144000
-
gel filtration
145000
-
gel filtration
150000
-
gel filtration
72000
77500
-
2 * 77500, SDS-PAGE
80000
-
x * 80000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
x-ray crystallography
?
-
x * 80000, SDS-PAGE
dimer
homodimer
-
-
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
enzyme in the apo, holo, and substrate-bound ternary forms, sitting drop vapor diffusion method, using 1.6 M Na/K-phosphate, 0.1 M HEPES pH 7.5 (apo form), or 30% (w/v) PEG3350, 0.1 M Bis-Tris pH 5.5, 0.3 M (NH4)2SO4 (holo form), or 20% (w/v) PEG3350, 0.1 M Bis-Tris pH 5.5, 0.1 M (NH4)2SO4 (ternary form)
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E392A
site-directed mutagenesis, kcat is reduced 12fold compared to the wild-type enzyme. The mutant shows no detectable adenosylcobalamin homolysis upon binding of the physiological substrate
E392D
site-directed mutagenesis, kcat is reduced 330fold compared to the wild-type enzyme. The mutant shows no detectable adenosylcobalamin homolysis upon binding of the physiological substrate
E392Q
site-directed mutagenesis, kcat is reduced 16fold compared to the wild-type enzyme. The mutant shows no detectable adenosylcobalamin homolysis upon binding of the physiological substrate
L692P
naturally occuring mutation, from patient with methylmalonic acidemia (MMA)
R108H
naturally occuring mutation, from patient with methylmalonic acidemia (MMA)
R228Q
naturally occuring mutation, from patient with methylmalonic acidemia (MMA)
R369H
naturally occuring mutation, from patient with methylmalonic acidemia (MMA)
G623R
-
six missense mutations, producing the amino acid changes G94V, Y231N, R369H, G623R, H678R and G717V are detected in L-methylmalonyl-CoA mutase cDNA of patients suffering from the mut-form of methylmalonic acidemia resulting from defective adenosylcobalamin binding. The mutations increase the Km for adenosylcobalamin by 40fold to 900fold, while the values for maximal velocity varies from 0.2% to nearly 100% of that of the wild-type protein
G717V
-
six missense mutations, producing the amino acid changes G94V, Y231N, R369H, G623R, H678R and G717V are detected in L-methylmalonyl-CoA mutase cDNA of patients suffering from the mut-form of methylmalonic acidemia resulting from defective adenosylcobalamin binding. The mutations increase the Km for adenosylcobalamin by 40fold to 900fold, while the values for maximal velocity varies from 0.2% to nearly 100% of that of the wild-type protein
G94V
-
six missense mutations, producing the amino acid changes G94V, Y231N, R369H, G623R, H678R and G717V are detected in L-methylmalonyl-CoA mutase cDNA of patients suffering from the mut-form of methylmalonic acidemia resulting from defective adenosylcobalamin binding. The mutations increase the Km for adenosylcobalamin by 40fold to 900fold, while the values for maximal velocity varies from 0.2% to nearly 100% of that of the wild-type protein
H678R
-
six missense mutations, producing the amino acid changes G94V, Y231N, R369H, G623R, H678R and G717V are detected in L-methylmalonyl-CoA mutase cDNA of patients suffering from the mut-form of methylmalonic acidemia resulting from defective adenosylcobalamin binding. The mutations increase the Km for adenosylcobalamin by 40fold to 900fold, while the values for maximal velocity varies from 0.2% to nearly 100% of that of the wild-type protein
R369H
-
six missense mutations, producing the amino acid changes G94V, Y231N, R369H, G623R, H678R and G717V are detected in L-methylmalonyl-CoA mutase cDNA of patients suffering from the mut-form of methylmalonic acidemia resulting from defective adenosylcobalamin binding. The mutations increase the Km for adenosylcobalamin by 40fold to 900fold, while the values for maximal velocity varies from 0.2% to nearly 100% of that of the wild-type protein
Y231N
-
six missense mutations, producing the amino acid changes G94V, Y231N, R369H, G623R, H678R and G717V are detected in L-methylmalonyl-CoA mutase cDNA of patients suffering from the mut-form of methylmalonic acidemia resulting from defective adenosylcobalamin binding. The mutations increase the Km for adenosylcobalamin by 40fold to 900fold, while the values for maximal velocity varies from 0.2% to nearly 100% of that of the wild-type protein
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
-
15 min, stable below
55
-
t1/2: 3 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
repeated freezing and thawing results in a gradual loss of activity
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 1.5 mg protein /ml in 50 mM potassium phosphate, pH 7.4, stable for at least 6 months
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni-NTA column chromatography, Resource Q column chromatography, and Superdex 200 gel filtration
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta(DE3)pLysS cells by nickel affinity and anion exchange chromatography
expressed in Saccharomyces cerevisiae
-
IMAC column chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3)R3-Rosetta cells
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta(DE3)pLysS cells
expression of the enzyme gene carrying a stop-codon mutation in mouse primary fibroblast cell lines, effects of gentamicin and PTC124 for stop-codon read-through potential, overview. Without treatment the cells contain 19% of the normal levels of methylmalonyl-CoA mutase enzyme activity which increases to 32% with treatment, suggesting a functional improvement. Treatment with PTC124 increases the amount of human methylmalonyl-CoA mutase gene mRNA by 1.6fold
functional transgenic enzyme expression in enzyme-deficient Mus musculus using a AAV8-CBA-MUT vector
expressed in Escherichia coli BL21(DE3) cells
-
mutant enzymes G94V, Y231N, R369H, G623R, H678R and G717V from patients suffering from the mut-form of methylmalonic acidemia, expression in Escherichia coli
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
establishment of a method to measure the concentration of succinyl-CoA with UPLC-MS/MS after enzyme reaction using peripheral lymphocytes, and investigation of the MCM enzyme activity of patients with methylmalonic acidemia
analysis
-
method for separation of methylmalonyl-CoA and succinyl-CoA by capillary electrophoresis suitable for evaluation of total and holo-enzyme activity in biological matrices. Application of method for the differential diagnosis of methylmalonic acidemia, in relation to protein or coenzyme defects
medicine
-
method for separation of methylmalonyl-CoA and succinyl-CoA by capillary electrophoresis suitable for evaluation of total and holo-enzyme activity in biological matrices. Application of method for the differential diagnosis of methylmalonic acidemia, in relation to protein or coenzyme defects
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Kolhouse, J.F.; Utley, C.; Allen, R.H.
Isolation and characterization of methylmalonyl-CoA mutase from human placenta
J. Biol. Chem.
255
2708-2712
1980
Homo sapiens
Manually annotated by BRENDA team
Fenton, W.A.; Hack, A.M.; Willard, H.F.; Gertler, A.; Rosenberg, L.E.
Purification and properties of methylmalonyl coenzyme A mutase from human liver
Arch. Biochem. Biophys.
214
815-823
1982
Homo sapiens
Manually annotated by BRENDA team
Ledley, F.D.; Lumetta, M.; Nguyen, P.N.; Kolhouse, J.F.; Allen, R.H.
Molecular cloning of L-methylmalonyl-CoA mutase: gene transfer and analysis of mut cell lines
Proc. Natl. Acad. Sci. USA
85
3518-3521
1988
Homo sapiens
Manually annotated by BRENDA team
Kolhouse, J.F.; Stabler, S.P.; Allen, R.H.
L-Methylmalonyl-CoA mutase from human placenta
Methods Enzymol.
166
407-414
1988
Homo sapiens
Manually annotated by BRENDA team
Taoka, S.; Padmakumar, R.; Lai, M.; Liu, H.; Banerjee, R.
Inhibition of the human methylmalonyl-CoA mutase by various CoA-esters
J. Biol. Chem.
269
31630-31634
1994
Homo sapiens
Manually annotated by BRENDA team
Janata, J.; Kogekar, N.; Fenton, W.A.
Expression and kinetic characterization of methylmalonyl-CoA mutase from patients with the mut-phenotype: evidence for naturally occuring interallelic complementation
Hum. Mol. Genet.
6
1457-1464
1997
Homo sapiens
Manually annotated by BRENDA team
Deodato, F.; Boenzi, S.; Santorelli, F.M.; Dionisi-Vici, C.
Methylmalonic and propionic aciduria
Am. J. Med. Genet. C Semin. Med. Genet.
142C
104-112
2006
Homo sapiens
Manually annotated by BRENDA team
Carlucci, F.; Rosi, F.; Tommassini, V.; Tabucchi, A.
CE assay of methylmalonyl-coenzyme-A mutase activity
Electrophoresis
28
1921-1925
2007
Homo sapiens, Rattus norvegicus
Manually annotated by BRENDA team
Takahashi-Iniguez, T.; Garcia-Arellano, H.; Trujillo-Roldan, M.A.; Flores, M.E.
Protection and reactivation of human methylmalonyl-CoA mutase by MMAA protein
Biochem. Biophys. Res. Commun.
404
443-447
2011
Homo sapiens
Manually annotated by BRENDA team
Froese, D.S.; Kochan, G.; Muniz, J.R.; Wu, X.; Gileadi, C.; Ugochukwu, E.; Krysztofinska, E.; Gravel, R.A.; Oppermann, U.; Yue, W.W.
Structures of the human GTPase MMAA and vitamin B12-dependent methylmalonyl-CoA mutase and insight into their complex formation
J. Biol. Chem.
285
38204-38213
2010
Homo sapiens (P22033), Homo sapiens
Manually annotated by BRENDA team
Buck, N.E.; Wood, L.R.; Hamilton, N.J.; Bennett, M.J.; Peters, H.L.
Treatment of a methylmalonyl-CoA mutase stopcodon mutation
Biochem. Biophys. Res. Commun.
427
753-757
2012
Homo sapiens (P22033), Homo sapiens
Manually annotated by BRENDA team
Makins, C.; Pickering, A.V.; Mariani, C.; Wolthers, K.R.
Mutagenesis of a conserved glutamate reveals the contribution of electrostatic energy to adenosylcobalamin co-C bond homolysis in ornithine 4,5-aminomutase and methylmalonyl-CoA mutase
Biochemistry
52
878-888
2013
Homo sapiens (P22033), Homo sapiens
Manually annotated by BRENDA team
Chandler, R.J.; Venditti, C.P.
Pre-clinical efficacy and dosing of an AAV8 vector expressing human methylmalonyl-CoA mutase in a murine model of methylmalonic acidemia (MMA)
Mol. Genet. Metab.
107
617-619
2012
Homo sapiens (P22033), Homo sapiens
Manually annotated by BRENDA team
Gotoh, K.; Nakajima, Y.; Tajima, G.; Hotta, Y.; Kataoka, T.; Kawade, Y.; Sugiyama, N.; Ito, T.; Kimura, K.; Maeda, Y.
Assay for methylmalonyl coenzyme A mutase activity based on determination of succinyl coenzyme A by ultrahigh-performance liquid chromatography tandem mass spectrometry
Anal. Bioanal. Chem.
407
5281-5286
2015
Homo sapiens (P22033), Homo sapiens
Manually annotated by BRENDA team
Takahashi-Iniguez, T.; Gonzalez-Noriega, A.; Michalak, C.; Flores, M.E.
Human MMAA induces the release of inactive cofactor and restores methylmalonyl-CoA mutase activity through their complex formation
Biochimie
142
191-196
2017
Homo sapiens (P22033), Homo sapiens
Manually annotated by BRENDA team