Information on EC 1.2.1.27 - methylmalonate-semialdehyde dehydrogenase (CoA-acylating)

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

EC NUMBER
COMMENTARY
1.2.1.27
-
RECOMMENDED NAME
GeneOntology No.
methylmalonate-semialdehyde dehydrogenase (CoA-acylating)
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2-methyl-3-oxopropanoate + CoA + H2O + NAD+ = propanoyl-CoA + HCO3- + NADH
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
oxidation
-
-
-
-
oxidation
Q6Z4E4
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
2,4-dinitrotoluene degradation
-
-
L-valine degradation I
-
-
Metabolic pathways
-
-
Propanoate metabolism
-
-
valine metabolism
-
-
Valine, leucine and isoleucine degradation
-
-
SYSTEMATIC NAME
IUBMB Comments
2-methyl-3-oxopropanoate:NAD+ 3-oxidoreductase (CoA-propanoylating)
Also converts 3-oxopropanoate into acetyl-CoA [3]. The reaction occurs in two steps with the decarboxylation process preceding CoA-binding [3]. Bicarbonate rather than CO2 is released as a final product [3].
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Aldh6a1
Q02252
gene name
methylmalonate semialdehyde dehydrogenase
-
-
methylmalonate semialdehyde dehydrogenase
P42412
-
methylmalonate semialdehyde dehydrogenase
Q6Z4E4
-
methylmalonate semialdehyde dehydrogenase
Q97YT9
-
methylmalonate semialdehyde dehydrogenase
Q97YT9
-
-
methylmalonate semialdehyde dehydrogenase protein
Q02252
-
methylmalonate-semialdehyde dehydrogenase
-
-
methylmalonate-semialdehyde dehydrogenase
Oryza sativa Japonica
-
-
-
methylmalonate-semialdehyde dehydrogenase
-
-
methylmalonic acid semialdehyde dehydrogenase
-
-
MMSDH
-
-
-
-
MMSDH
-
-
MMSDH
O49218
-
MMSDH
Oryza sativa Japonica
-
-
-
MSDH
P42412
-
MSDH
Q02252
-
MSDH
Q97YT9
-
-
OsALDH6
Q6Z4E4
-
SSO1218
Q97YT9
-
SSO1218
Q97YT9
-
-
CAS REGISTRY NUMBER
COMMENTARY
37205-49-5
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
Manually annotated by BRENDA team
Japonica
-
-
Manually annotated by BRENDA team
L. cv. Nipponbare
Swissprot
Manually annotated by BRENDA team
Oryza sativa Japonica
Japonica
-
-
Manually annotated by BRENDA team
Pseudomonas aeruginosa PAO
PAO
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
Q02252
MSDH malfunction can be a reason for 3-hydroxyisobutyric aciduria, which is a disorder of valine metabolism
metabolism
Q97YT9
for MSDH, a major function in the degradation of branched chain amino acids is proposed which is supported by the high sequence homology with characterized MSDHs from bacteria
metabolism
Q02252
MSDH enzyme is part of the catabolism pathway of L-valine
metabolism
-
for MSDH, a major function in the degradation of branched chain amino acids is proposed which is supported by the high sequence homology with characterized MSDHs from bacteria
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-methyl-3-oxopropanoate + CoA + H2O + NAD+
propanoyl-CoA + HCO3- + NADH
show the reaction diagram
Q97YT9
-
-
-
?
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
-
?
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
-
ir
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
-
ir
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
O49218
-
-
-
?, ir
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
-
ir
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
-
other acyl acceptor: 2-mercaptoethanol
-
ir
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
-
other acyl acceptor: 2-mercaptoethanol
-
ir
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
-
other acyl acceptor: 2-mercaptoethanol
-
ir
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
-
both stereoisomers are oxidized
-
ir
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
Pseudomonas aeruginosa PAO
-
-
-
-
ir
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
Pseudomonas aeruginosa PAO
-
other acyl acceptor: 2-mercaptoethanol
-
ir
2-methylbutyraldehyde + CoA + NAD+
?
show the reaction diagram
-
low activity
-
-
?
acetaldehyde + CoA + NAD+
?
show the reaction diagram
-
low activity
-
-
?
butyraldehyde + CoA + NAD+
?
show the reaction diagram
-
low activity
-
-
?
isobutyraldehyde + CoA + NAD+
?
show the reaction diagram
-
low activity
-
-
?
malonate semialdehyde + CoA + H2O + NAD+
acetyl-CoA + CO2 + NADH
show the reaction diagram
Q97YT9
-
-
-
?
malonate semialdehyde + CoA + NAD+
acetyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
-
?
malonate semialdehyde + CoA + NAD+
acetyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
-
-
malonate semialdehyde + CoA + NAD+
acetyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
ir
malonate semialdehyde + CoA + NAD+
acetyl-CoA + CO2 + NADH
show the reaction diagram
Q97YT9
-
-
-
?
malondialdehyde + CoA + NAD+
?
show the reaction diagram
-
low activity
-
-
?
methylmalonate semialdehyde + CoA + H2O + NAD+
propanoyl-CoA + HCO3- + NADH
show the reaction diagram
-
-
-
-
?
methylmalonate semialdehyde + CoA + H2O + NAD+
propanoyl-CoA + HCO3- + NADH
show the reaction diagram
Q02252
-
-
-
-
methylmalonate semialdehyde + CoA + H2O + NAD+
propanoyl-CoA + HCO3- + NADH
show the reaction diagram
Q97YT9
-
-
-
?
methylmalonate semialdehyde + CoA + H2O + NAD+
propanoyl-CoA + HCO3- + NADH
show the reaction diagram
P42412
weaker stabilization of the adenine ring triggers early NADH release in MSDH-catalyzed reaction
-
-
?
p-nitrophenyl acetate + CoA + NAD+
p-nitrophenol + acetyl-CoA + NADH
show the reaction diagram
-
-
-
ir
propanal + 2-mercaptoethanol + NAD+
? + NADH
show the reaction diagram
-
-
-
-
ir
propanal + 2-mercaptoethanol + NAD+
? + NADH
show the reaction diagram
-
-
-
ir
propanal + 2-mercaptoethanol + NAD+
? + NADH
show the reaction diagram
-
-
-
-
ir
propanal + 2-mercaptoethanol + NAD+
? + NADH
show the reaction diagram
Pseudomonas aeruginosa PAO
-
-
-
-
ir
propanal + CoA + NAD+
propanoyl-CoA + NADH
show the reaction diagram
-
-
-
-
ir
propanal + CoA + NAD+
propanoyl-CoA + NADH
show the reaction diagram
-
-
-
ir
propanal + NAD+
? + NADH
show the reaction diagram
-
-
-
?
propionaldehyde + CoA + H2O + NAD+
?
show the reaction diagram
Q97YT9
no physiological substrate
-
-
?
propionaldehyde + CoA + NAD+
? + NADH
show the reaction diagram
Q97YT9
highest activity, no physiological substrate for MSDH, it only forms the same thiopropionyl enzyme intermediate as methylmalonate semialdehyde and malonate semialdehyde
-
-
?
propionaldehyde + CoA + NAD+
propanoyl-CoA + NADH
show the reaction diagram
-
-
-
-
?
succinate semialdehyde + CoA + NAD+
?
show the reaction diagram
-
very low activity
-
-
?
methylmalonate-semialdehyde + CoA + NAD+
propionyl-CoA + NADH + H+ + HCO3-
show the reaction diagram
-
-
-
-
?
additional information
?
-
Q6Z4E4
ALDH superfamily represents a group of enzymes that catalyze the oxidation of endogenous and exogenous aldehydes to the corresponding carboxylic acids
-
-
-
additional information
?
-
Q97YT9
no activity is observed for the substrates D,L-glyceraldehyde, D,L-glyceraldehyde 3-phosphate and D,L-glycolaldehyde neither with NAD+ nor with NADP+ as cosubstrate
-
-
-
additional information
?
-
Q97YT9
no activity detected in the presence of NADP+ as cosubstrate, no activity with D,L-glyceraldehyde, D,L-glyceraldehyde-3-phosphate and D,L-glycolaldehyde - neither with NAD+ nor with NADP+ as cosubstrate
-
-
?
additional information
?
-
Q97YT9
no activity is observed for the substrates D,L-glyceraldehyde, D,L-glyceraldehyde 3-phosphate and D,L-glycolaldehyde neither with NAD+ nor with NADP+ as cosubstrate
-
-
-
additional information
?
-
Q97YT9
no activity detected in the presence of NADP+ as cosubstrate, no activity with D,L-glyceraldehyde, D,L-glyceraldehyde-3-phosphate and D,L-glycolaldehyde - neither with NAD+ nor with NADP+ as cosubstrate
-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2-methyl-3-oxopropanoate + CoA + H2O + NAD+
propanoyl-CoA + HCO3- + NADH
show the reaction diagram
Q97YT9
-
-
-
?
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
-
?
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
-
ir
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
-
ir
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
O49218
-
-
-
?
2-methyl-3-oxopropanoate + CoA + NAD+
propanoyl-CoA + CO2 + NADH
show the reaction diagram
Pseudomonas aeruginosa PAO
-
-
-
-
ir
malonate semialdehyde + CoA + NAD+
acetyl-CoA + CO2 + NADH
show the reaction diagram
-
-
-
-
ir
methylmalonate-semialdehyde + CoA + NAD+
propionyl-CoA + NADH + H+ + HCO3-
show the reaction diagram
-
-
-
-
?
additional information
?
-
Q6Z4E4
ALDH superfamily represents a group of enzymes that catalyze the oxidation of endogenous and exogenous aldehydes to the corresponding carboxylic acids
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
CoA
Q97YT9
enzyme is strictly CoA-dependent
NAD+
Q6Z4E4
-
NAD+
Q97YT9
cosubstrate; no activity in the presence of NADP+ as cosubstrate
NADP+
Q6Z4E4
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NADH
-
competitive with NAD+
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.54
2-mercaptoethanol
-
-
44
2-mercaptoethanol
-
-
0.0025
2-Methyl-3-oxopropanoate
-
-
0.0053
2-Methyl-3-oxopropanoate
-
-
0.019
2-Methyl-3-oxopropanoate
-
-
0.024
2-Methyl-3-oxopropanoate
-
-
0.06
2-Methyl-3-oxopropanoate
-
50 mM KP04 pH 8.2, 30C, 0.5 mM 2-methyl-3-oxopropanoate, 12 mM NAD+, 0.5 mM CoA
0.03
CoA
-
-
0.063
CoA
P42412
V229G/G225 insertion, pH 8.2 and 30C
0.085
CoA
-
wild type MSDH, steady-state conditions with propionaldehyde, pH 8.2 and 30C
0.096
CoA
P42412
V229G/H226P/G225 insertion, pH 8.2 and 30C
0.119
CoA
P42412
V229G/H226P, pH 8.2 and 30C
0.12
CoA
-
50 mM potassium phosphate pH 8.2, 30C, 0.5 mM 2-methyl-3-oxopropanoate, 12 mM NAD+, 0.5 mM CoA
0.12
CoA
-
wild type MSDH, steady-state conditions with methylmalonate semialdehyde, pH 8.2 and 30C
0.12
CoA
P42412
wild type MSDH, pH 8.2 and 30C
0.151
CoA
P42412
G225 insertion, pH 8.2 and 30C
0.234
CoA
P42412
V229G, pH 8.2 and 30C
0.33
CoA
-
R124L mutant enzyme, steady-state conditions with propionaldehyde, pH 8.2 and 30C
0.497
CoA
P42412
V229G/Y252L/V253I/G225 insertion, pH 8.2 and 30C
0.57
CoA
-
R124L mutant enzyme, steady-state conditions with methylmalonate semialdehyde, pH 8.2 and 30C
0.62
CoA
-
R301L mutant enzyme, steady-state conditions with methylmalonate semialdehyde, pH 8.2 and 30C
0.0045
Malonate semialdehyde
-
-
0.006
methylmalonate semialdehyde
-
R124L mutant enzyme, steady-state conditions, pH 8.2 and 30C
0.021
methylmalonate semialdehyde
P42412
V229G/H226P/G225 insertion, pH 8.2 and 30C
0.022
methylmalonate semialdehyde
P42412
G225 insertion, pH 8.2 and 30C
0.023
methylmalonate semialdehyde
-
R301L mutant enzyme, steady-state conditions, pH 8.2 and 30C
0.027
methylmalonate semialdehyde
P42412
V229G/G225 insertion, pH 8.2 and 30C
0.028
methylmalonate semialdehyde
P42412
V229G/H226P, pH 8.2 and 30C
0.054
methylmalonate semialdehyde
P42412
V229G, pH 8.2 and 30C
0.06
methylmalonate semialdehyde
-
wild type MSDH, steady-state conditions, pH 8.2 and 30C
0.06
methylmalonate semialdehyde
P42412
wild type MSDH, pH 8.2 and 30C
0.033
NAD+
-
wild type MSDH, steady-state conditions with propionaldehyde, pH 8.2 and 30C
0.038
NAD+
-
R124L mutant enzyme, steady-state conditions with propionaldehyde, pH 8.2 and 30C
0.06
NAD+
-
R124L mutant enzyme, steady-state conditions with methylmalonate semialdehyde, pH 8.2 and 30C
0.12
NAD+
-
R301L mutant enzyme, steady-state conditions with methylmalonate semialdehyde, pH 8.2 and 30C
0.15
NAD+
-
-
0.15
NAD+
P42412
V229G/G225 insertion, pH 8.2 and 30C
0.57
NAD+
P42412
V229G, pH 8.2 and 30C
0.66
NAD+
P42412
V229G/H226P, pH 8.2 and 30C
0.69
NAD+
P42412
G225 insertion, pH 8.2 and 30C
0.77
NAD+
P42412
V229G/H226P/G225 insertion, pH 8.2 and 30C
2.3
NAD+
-
50 mM potassium phosphate pH 8.2, 30C, 0.5 mM 2-methyl-3-oxopropanoate, 12 mM NAD+, 0.5 mM CoA
2.3
NAD+
-
wild type MSDH, steady-state conditions with methylmalonate semialdehyde, pH 8.2 and 30C
2.3
NAD+
P42412
wild type MSDH, pH 8.2 and 30C
9.44
NAD+
P42412
V229G/Y252L/V253I/G225 insertion, pH 8.2 and 30C
6.1
propanal
-
-
6.4
propanal
-
-
9.3
propionaldehyde
-
R124L mutant enzyme, steady-state conditions, pH 8.2 and 30C
9.8
propionaldehyde
-
wild type MSDH, steady-state conditions, pH 8.2 and 30C
0.215
methylmalonate semialdehyde
P42412
V229G/Y252L/V253I/G225 insertion, pH 8.2 and 30C
additional information
additional information
-
Km-value of CoA for R301L mutant enzyme under steady-state conditions with propionaldehyde is above 1 mM at pH 8.2 and 30C
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.003
2-Methyl-3-oxopropanoate
Q97YT9
pH 8.2, 30C
1.1
2-Methyl-3-oxopropanoate
-
50 mM potassium phosphate pH 8.2, 30C, 0.5 mM 2-methyl-3-oxopropanoate, 12 mM NAD+, 0.5 mM CoA
2.8
CoA
-
R124L mutant enzyme, steady-state conditions with propionaldehyde, pH 8.2 and 30C
12.6
CoA
-
wild type MSDH, steady-state conditions with propionaldehyde, pH 8.2 and 30C
0.02
Malonate semialdehyde
Q97YT9
30C, pH 8.2; pH 8.2, 30C
0.02
NAD+
P42412
V229G/H226P/G225 insertion, pH 8.2 and 30C; V229G/Y252L/V253I/G225 insertion, pH 8.2 and 30C
0.06
NAD+
-
R124L mutant enzyme, steady-state conditions with methylmalonate semialdehyde, pH 8.2 and 30C
0.09
NAD+
P42412
V229G/H226P, pH 8.2 and 30C
0.14
NAD+
-
R301L mutant enzyme, steady-state conditions with methylmalonate semialdehyde, pH 8.2 and 30C
0.21
NAD+
P42412
V229G/G225 insertion, pH 8.2 and 30C
0.39
NAD+
P42412
G225 insertion, pH 8.2 and 30C
0.78
NAD+
P42412
V229G, pH 8.2 and 30C
2.2
NAD+
-
wild type MSDH, steady-state conditions with methylmalonate semialdehyde, pH 8.2 and 30C
2.2
NAD+
P42412
wild type MSDH, pH 8.2 and 30C
0.03
propionaldehyde
Q97YT9
30C, pH 8.2; pH 8.2, 30C
0.003
methylmalonate semialdehyde
Q97YT9
30C, pH 8.2
additional information
additional information
-
Kcat-value of CoA for R301L mutant enzyme under steady-state conditions with propionaldehyde is above 3 1/sec at pH 8.2 and 30C
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3
CoA
-
R301L mutant enzyme, steady-state conditions with propionaldehyde, pH 8.2 and 30C
18
8.5
CoA
-
R124L mutant enzyme, steady-state conditions with propionaldehyde, pH 8.2 and 30C
18
150
CoA
-
wild type MSDH, steady-state conditions with propionaldehyde, pH 8.2 and 30C
18
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0031
NADH
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8
-
2-methyl-3-oxopropanoate + CoA + NAD+
8.5
-
2-methyl-3-oxopropanoate + 2-mercaptoethanol + NAD+; 2-methyl-3-oxopropanoate + CoA + NAD+; propanal + 2-mercaptoethanol + NAD+
9
-
propanal + NAD+
9
-
propanal + CoA + NAD+
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7 - 10
-
methylmalonate semialdehyde oxidation, more than 90% of maximal activity
7 - 8.5
-
methylmalonate semialdehyde oxidation, less than 90% of maximal activity above and below
8 - 10
-
propanal oxidation, less than 50% of maximal activity below, less than 60% of maximal activity above
8 - 9.5
-
propanal oxidation, less than 50% of maximal activity above and below
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6
O49218
2D-PAGE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
mRNA expression
Manually annotated by BRENDA team
O49218
sheats
Manually annotated by BRENDA team
Oryza sativa Japonica
-
mRNA expression
-
Manually annotated by BRENDA team
O49218
MMSDH expression is induced in auxin-stimulated and zinc-stimulated root formation
Manually annotated by BRENDA team
-
enhanced mRNA expression after exposure to gibberellin
Manually annotated by BRENDA team
Oryza sativa Japonica
-
enhanced mRNA expression after exposure to gibberellin
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Oryza sativa Japonica
-
-
-
Manually annotated by BRENDA team
-
mitochondrial targeting sequence
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Bacillus subtilis (strain 168)
Rhizobium meliloti (strain 1021)
Rhizobium meliloti (strain 1021)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50000
Q97YT9
SDS-PAGE
722211
54600
Q97YT9
calculated from sequence
722211
57200
Q6Z4E4
-
698095
130000
Q97YT9
native molecular weight using gel filtration
722211
132000
-
gel filtration
390219
250000
-
gel filtration
288055
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
O49218
x * 53700, 2D-PAGE
dimer
-
2 * 58000, SDS-PAGE, 2 * 69000, gel filtration in presence of SDS
homodimer
Q97YT9
2 * 50000
homodimer
-
2 * 50000
-
tetramer
-
4 * 58000, SDS-PAGE
tetramer
-
4 * 55330, cDNA sequence
tetramer
P42412
dimer of dimers, monomer consists of three domains, the dinucleotide binding domain comprising the residues 3123 and 141251, the catalytic domain with residues 252270, and a small domain, with residues 124140 and 471486
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
phosphoprotein
-
-
side-chain modification
-
acylation of active site cysteine residue Cys-285 by long-chain fatty acyl-CoA esters inactivates the enzyme
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging-drop vapour diffusion, crystals diffract to 2.5 A resolution
-
methylmalonate semialdehyde dehydrogenase in binary complex with NAD+, showing that the nicotinamide ring is well defined in the electron density due to direct and H2O-mediated hydrogen bonds with the carboxamide and that a conformational isomerization of the NMNH is possible in MSDH
P42412
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
on ice, 1 mM NAD+, 20 h, 77% activity remains
-
on ice, 10% glycerol, 20 h, 32% activity remains
-
on ice, 20 h, 11% activity remains
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
MSDH purified by ammonium sulfate fractionation, 40-80%, and gel filtration on a ACA 34 resin
-
recombinant MSDH
-
; to apparent homogeneity using heat precipitation at 80C for 20 min, ion exchange chromatography and gel filtration
Q97YT9
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
-
in Escherichia coli DH5[alpha] transformants containing the pSKmsdbsub plasmid
P42412
into the pGEM-T easy vector for sequencing
Q6Z4E4
expression in Escherichia coli; SSO1218 expressed in Escherichia coli BL21 (DE3) RIL via isopropyl-b-D-thiogalactopyranoside induction
Q97YT9
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
N427L
P42412
substitution dramatically alters the catalytic properties of the enzyme, acylation becomes rate-limiting with a decrease of the associated rate constant by at least 10000fold relative to the wild-type MSDH
R124L
-
results obtained under pre-steady conditions show that both Arg residues participate not only in methylmalonate semialdehyde binding via stabilizing interactions between the guanidinium groups and the carboxylate but also in the formation of an efficient MSDH-NAD+-methylmalonate semialdehyde ternary complex
R301L
-
results obtained under pre-steady conditions show that both Arg residues participate not only in methylmalonate semialdehyde binding via stabilizing interactions between the guanidinium groups and the carboxylate but also in the formation of an efficient MSDH-NAD+-methylmalonate semialdehyde ternary complex
V229G
P42412
significant decrease of the rate constant associated with the dissociation of NADH from the NADH-thioacylenzyme complex
V229G
P42412
together with insertion of glycine at position 225, significant decrease of the rate constant associated with the dissociation of NADH from the NADH/thioacylenzyme complex
V229G/H226P
P42412
rate-limiting step changes to acylation instead of deacylation
V229G/H226P
P42412
together with insertion of glycine at position 225, rate-limiting step changes to acylation instead of deacylation
V229G/Y252L/V253I
P42412
together with insertion of glycine at position 225, rate-limiting step changes to acylation instead of deacylation
W177F
-
NAD-binding followed by fluorescence quenching of methylmalonate semialdehyde dehydrogenase. Only W468 is responsible for time-dependent additional quenching
W28F
-
NAD-binding followed by fluorescence quenching of methylmalonate semialdehyde dehydrogenase. Only W468 is responsible for time-dependent additional quenching
W397F
-
NAD-binding followed by fluorescence quenching of methylmalonate semialdehyde dehydrogenase. Only W468 is responsible for time-dependent additional quenching
W468F
-
NAD-binding followed by fluorescence quenching of methylmalonate semialdehyde dehydrogenase. Only W468 is responsible for time-dependent additional quenching
W76F
-
NAD-binding followed by fluorescence quenching of methylmalonate semialdehyde dehydrogenase. Only W468 is responsible for time-dependent additional quenching
P62S
Q02252
missense mutation in a highly conserved amino acid of MSDH, patient has severe developmental delay associated with development of marked post-natal microcephaly, at 7 years static moderate learning difficulties and borderline microcephaly
S262Y
Q02252
missense mutation in a highly conserved amino acid of MSDH, patient developed a febrile illness and died from a hepatoencephalopathy at 2 years of age
C285A
-
expressed in Escherichia coli, no activity
N251E
-
expressed in Escherichia coli, no activity
C49A/C176A/C305A/C369A/C403A
-
kinetic parameters similar to the wild-type enzmye
additional information
P42412
further mutant enzyme created by single insertion of glycine at position 225 resulting in significant decrease of the rate constant associated with the dissociation of NADH fromthe NADH/thioacylenzyme complex