Cloned (Comment) | Organism |
---|---|
recombinant expression of C-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
R227P | a naturally occuring GCDH disease-related mutation involved in glutaric aciduria type I (GA-I). Heterozygous patients harbouring the two mutations GCDH-p.Arg227Pro and GCDH-p.Val400Met show increased residual enzymatic activity in relation to homozygous patients with only one of the mutations, suggesting a complementation effect between the two. The mutant shows 95% reduced activity compared to wild-type | Homo sapiens |
V400M | a naturally occuring GCDH disease-related mutation involved in glutaric aciduria type I (GA-I). Heterozygous patients harbouring the two mutations GCDH-p.Arg227Pro and GCDH-p.Val400Met show increased residual enzymatic activity in relation to homozygous patients with only one of the mutations, suggesting a complementation effect between the two. Thermal stress affects cofactor binding in the GCDH-p.Val400Met mutant. In vivo the p.Val400Met variant displays impaired interaction with the partner ETF, resulting in the lower values observed in patient fibroblasts. The mutant shows 24% reduced activity compared to wild-type | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten steady-state kinetic analysis for wild-type and mutant enzymes | Homo sapiens | |
0.02 | - |
glutaryl-CoA | recombinant mutant V400M with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens | |
0.0211 | - |
glutaryl-CoA | recombinant wild-type enzyme with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens | |
0.0326 | - |
glutaryl-CoA | recombinant mutant R227P with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | - |
Homo sapiens | 5739 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
glutaryl-CoA + electron-transfer flavoprotein | Homo sapiens | overall reaction | crotonyl-CoA + CO2 + reduced electron-transfer flavoprotein | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q92947 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Homo sapiens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
5 | - |
recombinant mutant R227P with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens |
70 | - |
recombinant mutant V400M with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens |
112 | - |
recombinant mutant R227P with PMS, pH 7.8, temperature not specified in the publication | Homo sapiens |
205 | - |
recombinant wild-type enzyme with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens |
1683 | - |
recombinant mutant V400M with PMS, pH 7.8, temperature not specified in the publication | Homo sapiens |
2224 | - |
recombinant wild-type enzyme with PMS, pH 7.8, temperature not specified in the publication | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
glutaryl-CoA + electron-transfer flavoprotein | overall reaction | Homo sapiens | crotonyl-CoA + CO2 + reduced electron-transfer flavoprotein | - |
? | |
additional information | GCDH enzyme activity is measured following the reduction of DCPIP at 600 nm, with either PMS (artificial electron acceptor) or recombinant human ETF (GCDH natural electron acceptor), and glutaryl-CoA as electron donor, in 10 mM HEPES, pH 7.8 | Homo sapiens | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
GCDH | - |
Homo sapiens |
glutaryl-CoA dehydrogenase | - |
Homo sapiens |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
43 | 53 | apparent thermal denaturation midpoint (Tm) of GCDH wild -type and mutants using different techniques, overview | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.6 | - |
glutaryl-CoA | recombinant mutant R227P with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens | |
1.3 | - |
glutaryl-CoA | recombinant mutant V400M with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens | |
3.1 | - |
glutaryl-CoA | recombinant wild-type enzyme with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.8 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
electron transferring flavoprotein | ETF, the GCDH natural electron acceptor | Homo sapiens | |
FAD | wild-type and mutant FAD binding structures, overview | Homo sapiens | |
additional information | kinetics of flavin release, overview | Homo sapiens | |
phenazine methosulfate | PMS, an artificial electron acceptor | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
malfunction | glutaric aciduria type I (GA-I) is an autosomal recessive neurometabolic disease caused by mutations in the GCDH gene that encodes for glutaryl-CoA dehydrogenase (GCDH), a flavoprotein involved in the metabolism of tryptophan, lysine and hydroxylysin | Homo sapiens |
metabolism | glutaryl-CoA dehydrogenase (GCDH), a flavoprotein, is involved in the metabolism of tryptophan, lysine and hydroxylysin | Homo sapiens |
additional information | structure comparisons of wild-type and mutant enzyme proteins, FAD binding structures, overview | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
18.4 | - |
glutaryl-CoA | recombinant mutant R227P with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens | |
65 | - |
glutaryl-CoA | recombinant mutant V400M with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens | |
146.92 | - |
glutaryl-CoA | recombinant wild-type enzyme with ETF, pH 7.8, temperature not specified in the publication | Homo sapiens |