EC Number | Cloned (Comment) | Organism |
---|---|---|
1.2.4.4 | expression of N-and C-terminally His6-tagged component E1b and mutants, and of C-terminally His-tagged lipoic acid-bearing domain, amino acid residues 1-84 of component E2b, in vitro lipoylation of the latter | Homo sapiens |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.2.4.4 | purified wild-type and C-terminally His-tagged recombinant E1b component and mutants, 22°C, hanging drop vapour diffusion method, Mg2+ or Mn2+, X-ray diffraction structure determination and analysis at 1.8 A resolution | Homo sapiens |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.2.4.4 | D295A | site-directed mutagenesis of an alpha-subunit residue of component E1b, increased activity compared to the wild-type | Homo sapiens |
1.2.4.4 | R287A | site-directed mutagenesis of an alpha-subunit residue of component E1b, highly increased Km and reduced activity compared to the wild-type | Homo sapiens |
1.2.4.4 | R301A | site-directed mutagenesis of an alpha-subunit residue of component E1b, increased Km and reduced activity compared to the wild-type | Homo sapiens |
1.2.4.4 | Y300A | site-directed mutagenesis of an alpha-subunit residue of component E1b, increased Km and reduced activity compared to the wild-type | Homo sapiens |
1.2.4.4 | Y300F | site-directed mutagenesis of an alpha-subunit residue of component E1b, increased Km and reduced activity compared to the wild-type | Homo sapiens |
4.1.1.72 | D295A | of component E1b, 2fold increase in kcat-value | Homo sapiens |
4.1.1.72 | R287A | of component E1b, 40fold increase in Km-value for 2-oxoisovalerate | Homo sapiens |
4.1.1.72 | R301A | of component E1b, 4fold increase in Km-value for 2-oxoisovalerate | Homo sapiens |
4.1.1.72 | Y300A | of component E1b, 6fold increase in Km-value for 2-oxoisovalerate | Homo sapiens |
4.1.1.72 | Y300F | of component E1b, 5fold increase in Km-value for 2-oxoisovalerate | Homo sapiens |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
2.7.11.4 | additional information | binding of thiamin diphosphate cofactor to branched-chain alpha-keto acid decarboxylase/dehydrogenase, which induces a phosphorylation loop conformation change, inhibits the phosphorylation of the protein by the BCKD kinase, no inhibition of phosphorylation of mutant R287A, D295A, Y300F, and R301A E1B components | Rattus norvegicus |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.2.4.4 | additional information | - |
additional information | kinetics of the overall reaction, thermodynamics and dissociation constants of interaction between component E1b and the lipoic acid-bearing domain of component E2b | Homo sapiens | |
1.2.4.4 | 0.007 | - |
thiamine diphosphate | recombinant wild-type component E1b, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.017 | - |
thiamine diphosphate | recombinant component E1b mutant D295Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.024 | - |
thiamine diphosphate | recombinant component E1b mutant R287Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.029 | - |
thiamine diphosphate | recombinant component E1b mutants R301Aalpha and Y300Falpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.04 | - |
thiamine diphosphate | recombinant component E1b mutant Y300Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.05 | - |
2-oxo-isovalerate | recombinant wild-type component E1b, and mutant D295Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.23 | - |
2-oxo-isovalerate | recombinant component E1b mutant R301Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.25 | - |
2-oxo-isovalerate | recombinant component E1b mutant Y300Falpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.34 | - |
2-oxo-isovalerate | recombinant component E1b mutant Y300Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 2.33 | - |
2-oxo-isovalerate | recombinant component E1b mutant R287Aalpha, pH 7.5, 22°C | Homo sapiens | |
4.1.1.72 | 0.00063 | - |
thiamine diphosphate | wild-type, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.0066 | - |
thiamine diphosphate | wild-type, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.015 | - |
thiamine diphosphate | mutant D295A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.016 | - |
thiamine diphosphate | mutant D295A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.018 | - |
thiamine diphosphate | mutant Y300F, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.026 | - |
thiamine diphosphate | mutant R301A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.026 | - |
thiamine diphosphate | mutant Y300A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.028 | - |
thiamine diphosphate | mutant Y300F, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.029 | - |
thiamine diphosphate | mutant R301A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.04 | - |
thiamine diphosphate | mutant Y300A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.05 | - |
2-oxoisovalerate | mutant D295A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.05 | - |
2-oxoisovalerate | mutant D295A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.05 | - |
2-oxoisovalerate | wild-type, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.05 | - |
2-oxoisovalerate | wild-type, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.14 | - |
2-oxoisovalerate | mutant Y300A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.225 | - |
2-oxoisovalerate | mutant R301A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.252 | - |
2-oxoisovalerate | mutant Y300F, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.255 | - |
2-oxoisovalerate | mutant R301A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.337 | - |
2-oxoisovalerate | mutant Y300A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.39 | - |
2-oxoisovalerate | mutant Y300F, pH 7.5, 30°C, overall reaction | Homo sapiens |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.2.4.4 | Mg2+ | required for the binding of cofactor thiamine diphosphate to component E1b, Mn2+ can substitute | Homo sapiens | |
2.7.11.4 | Mg2+ | - |
Rattus norvegicus |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.2.4.4 | 3-methyl-2-oxobutanoate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine | Homo sapiens | - |
[dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] S-(2-methylpropanoyl)dihydrolipoyllysine + CO2 | - |
? | |
2.7.11.4 | ATP + branched-chain alpha-keto acid decarboxylase/dehydrogenase | Rattus norvegicus | inactivation of the substrate enzyme | ADP + phosphorylated branched-chain alpha-keto acid decarboxylase/dehydrogenase | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.2.4.4 | Homo sapiens | - |
- |
- |
2.7.11.4 | Rattus norvegicus | - |
- |
- |
4.1.1.72 | Homo sapiens | - |
- |
- |
EC Number | Posttranslational Modification | Comment | Organism |
---|---|---|---|
1.2.4.4 | additional information | limited proteolytic analysis of E1b component, overview | Homo sapiens |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
1.2.4.4 | 3-methyl-2-oxobutanoate + [dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] lipoyllysine = [dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] S-(2-methylpropanoyl)dihydrolipoyllysine + CO2 + 2 H+ | substrate recognition and reaction mechanism | Homo sapiens | |
4.1.1.72 | (3S)-3-methyl-2-oxopentanoate = 2-methylbutanal + CO2 | binding of cofactor thiamine diphosphate to E1b component of enzyme induces a disorder-to-order transition of the conserved phosphorylation loop carrying phosphorylation sites S292 and S302 | Homo sapiens |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
1.2.4.4 | additional information | - |
decarboxylation activity, 30°C | Homo sapiens |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.2.4.4 | 2-oxoisovalerate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine | - |
Homo sapiens | ? + CO2 | - |
? | |
1.2.4.4 | 3-methyl-2-oxobutanoate + [dihydrolipoyllysine-residue(2-methylpropanoyl)transferase]-lipoyllysine | - |
Homo sapiens | [dihydrolipoyllysine-residue (2-methylpropanoyl)transferase] S-(2-methylpropanoyl)dihydrolipoyllysine + CO2 | - |
? | |
1.2.4.4 | additional information | the overall reaction proceeds in several steps of the components E1, E2, and E3, overview, conformation of the conserved phosphorylation loop, carrying 2 phosphorylation sites Ser292alpha andSer302alpha, is essential for recognition of lipoylated LBD to initiate E1b-catalyzed reductive acylation, E1b is regulated by reversible phosphorylation through the kinase of the multienzyme complex | Homo sapiens | ? | - |
? | |
2.7.11.4 | ATP + branched-chain alpha-keto acid decarboxylase/dehydrogenase | inactivation of the substrate enzyme | Rattus norvegicus | ADP + phosphorylated branched-chain alpha-keto acid decarboxylase/dehydrogenase | - |
? | |
2.7.11.4 | ATP + branched-chain alpha-keto acid decarboxylase/dehydrogenase | recombinant human wild-type and mutant substrate proteins, overview, phosphorylation at Ser301 and Ser302 in the phosphorylation loop of decarboxylase E1b component of the large branched-chain alpha-keto acid dehydrogenase complex, loop structure, overview | Rattus norvegicus | ADP + phosphorylated branched-chain alpha-keto acid decarboxylase/dehydrogenase | - |
? | |
4.1.1.72 | 2-oxoisovalerate | - |
Homo sapiens | 2-methylpropanal + CO2 | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.2.4.4 | BCKD | - |
Homo sapiens |
1.2.4.4 | branched-chain alpha-keto acid decarboxylase/dehydrogenase | - |
Homo sapiens |
2.7.11.4 | BCKD kinase | - |
Rattus norvegicus |
2.7.11.4 | branched-chain alpha-keto acid decarboxylase/dehydrogenase kinase | - |
Rattus norvegicus |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
2.7.11.4 | 22 | - |
assay at room temperature | Rattus norvegicus |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.2.4.4 | 0.195 | - |
2-oxo-isovalerate | recombinant wild-type component E1b, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.197 | - |
thiamine diphosphate | recombinant wild-type component E1b, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.31 | - |
thiamine diphosphate | recombinant component E1b mutant Y300Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.36 | - |
2-oxo-isovalerate | recombinant component E1b mutant Y300Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.38 | - |
2-oxo-isovalerate | recombinant component E1b mutant R287Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.39 | - |
thiamine diphosphate | recombinant component E1b mutant R287Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.41 | - |
2-oxo-isovalerate | recombinant component E1b mutant R301Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.42 | - |
thiamine diphosphate | recombinant component E1b mutants R301Aalpha and D295Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.485 | - |
2-oxo-isovalerate | recombinant component E1b mutant D295Aalpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.53 | - |
thiamine diphosphate | recombinant component E1b mutant Y300Falpha, pH 7.5, 22°C | Homo sapiens | |
1.2.4.4 | 0.57 | - |
2-oxo-isovalerate | recombinant component E1b mutant Y300Falpha, pH 7.5, 22°C | Homo sapiens | |
4.1.1.72 | 0.19 | - |
thiamine diphosphate | wild-type, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.19 | - |
2-oxoisovalerate | wild-type, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.19 | - |
thiamine diphosphate | wild-type, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.19 | - |
2-oxoisovalerate | wild-type, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.31 | - |
thiamine diphosphate | mutant Y300A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.31 | - |
thiamine diphosphate | mutant Y300A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.38 | - |
2-oxoisovalerate | mutant Y300A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.38 | - |
2-oxoisovalerate | mutant Y300A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.41 | - |
thiamine diphosphate | mutant R301A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.41 | - |
2-oxoisovalerate | mutant R301A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.41 | - |
thiamine diphosphate | mutant R301A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.41 | - |
2-oxoisovalerate | mutant R301A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.42 | - |
thiamine diphosphate | mutant D295A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.42 | - |
thiamine diphosphate | mutant D295A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.48 | - |
2-oxoisovalerate | mutant D295A, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.48 | - |
2-oxoisovalerate | mutant D295A, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.53 | - |
thiamine diphosphate | mutant Y300F, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.53 | - |
thiamine diphosphate | mutant Y300F, pH 7.5, 30°C, overall reaction | Homo sapiens | |
4.1.1.72 | 0.57 | - |
2-oxoisovalerate | mutant Y300F, pH 7.5, 30°C, E1b-catalyzed decarboxylation reaction | Homo sapiens | |
4.1.1.72 | 0.57 | - |
2-oxoisovalerate | mutant Y300F, pH 7.5, 30°C, overall reaction | Homo sapiens |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.2.4.4 | 7.5 | - |
assay at | Homo sapiens |
2.7.11.4 | 7.4 | - |
assay at | Rattus norvegicus |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.2.4.4 | thiamine diphosphate | bound to component E1, heterotetrameric cofactor binding fold, cofactor binding prevents phosphorylation of E1b and inactivates it by inducing a disorder-to-order transition of the conserved phosphorylation loop carrying 2 phosphorylation sites Ser292alpha andSer302alpha, cross-talk between thiamine diphosphate and the phosphorylation loop conformation as a feed-forward switch for th complex reaction | Homo sapiens | |
2.7.11.4 | ATP | - |
Rattus norvegicus | |
4.1.1.72 | thiamine diphosphate | binds to E1b component of enzyme complex | Homo sapiens |