Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and beta-subunit mutant enzymes | Acetoanaerobium sticklandii |
Protein Variants | Comment | Organism |
---|---|---|
C700S | site-directed mutagenesis, the beta-subunit mutant shows similar kinetics and activity as the wild-type enzyme | Acetoanaerobium sticklandii |
D627A | site-directed mutagenesis, the beta-subunit mutant shows similar kinetics and slightly reduced activity compared to the wild-type enzyme | Acetoanaerobium sticklandii |
E338A | site-directed mutagenesis, the beta-subunit mutant shows altered kinetics and highly reduced activity compared to the wild-type enzyme | Acetoanaerobium sticklandii |
G128D | site-directed mutagenesis, inactive beta-subunit mutant | Acetoanaerobium sticklandii |
G339W | site-directed mutagenesis, the beta-subunit mutant shows altered kinetics and highly reduced activity compared to the wild-type enzyme | Acetoanaerobium sticklandii |
I424E | site-directed mutagenesis, the beta-subunit mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Acetoanaerobium sticklandii |
additional information | OAM variants are designed to perturb the interface between the cobalamin-binding domain and the pyridoxal 5'-phosphate-binding TIM-barrel domain. Steady-state and single turnover kinetic studies of these variants, combined with pulsed electron-electron double resonance measurements of spin-labeled OAM are used to provide direct evidence for a dynamic interface between the cobalamin and pyridoxal 5'-phosphate-binding domains, overview | Acetoanaerobium sticklandii |
P343W | site-directed mutagenesis, the beta-subunit mutant shows altered kinetics and highly reduced activity compared to the wild-type enzyme | Acetoanaerobium sticklandii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state and pre-steady-state kinetics | Acetoanaerobium sticklandii | |
0.176 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant E338A | Acetoanaerobium sticklandii | |
0.185 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant I424E | Acetoanaerobium sticklandii | |
0.186 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant C700S | Acetoanaerobium sticklandii | |
0.189 | - |
D-ornithine | pH 8.5, 25°C, recombinant wild-type enzyme | Acetoanaerobium sticklandii | |
0.19 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant G339W | Acetoanaerobium sticklandii | |
0.193 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant D627A | Acetoanaerobium sticklandii | |
0.193 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant P343W | Acetoanaerobium sticklandii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-ornithine | Acetoanaerobium sticklandii | - |
(2R,4S)-2,4-diaminopentanoate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acetoanaerobium sticklandii | E3PY95 AND E3PY96 | subunits beta and alpha | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and beta-subunit mutant enzymes | Acetoanaerobium sticklandii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-ornithine | - |
Acetoanaerobium sticklandii | (2R,4S)-2,4-diaminopentanoate | - |
r | |
additional information | cobalamin-dependent enzymes enhance the rate of C-Co bond cleavage by up to 1012-fold to generate cob(II)alamin and a transient adenosyl radical. In the case of the pyridoxal 5'-phosphate and cobalamin-dependent enzymes lysine 5,6-aminomutase (EC 5.4.3.3) and ornithine 4,5 aminomutase, it has been proposed that a large scale domain reorientation of the cobalamin-binding domain is linked to radical catalysis Coupled enzyme assay with (2R,4S)-2,4-diaminopentanoate dehydrogenase (DAPDH) from Clostridium difficile | Acetoanaerobium sticklandii | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
OAM | - |
Acetoanaerobium sticklandii |
ornithine 4,5-aminomutase | - |
Acetoanaerobium sticklandii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Acetoanaerobium sticklandii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.14 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant P343W | Acetoanaerobium sticklandii | |
0.2 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant G339W | Acetoanaerobium sticklandii | |
0.24 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant E338A | Acetoanaerobium sticklandii | |
0.76 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant I424E | Acetoanaerobium sticklandii | |
2.88 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant C700S | Acetoanaerobium sticklandii | |
2.89 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant D627A | Acetoanaerobium sticklandii | |
2.97 | - |
D-ornithine | pH 8.5, 25°C, recombinant wild-type enzyme | Acetoanaerobium sticklandii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Acetoanaerobium sticklandii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
5'-deoxyadenosylcobalamin | - |
Acetoanaerobium sticklandii | |
additional information | ornithine 4,5-aminomutase is a 5'-deoxyadenosylcobalamin and pyridoxal 5'-phosphate co-dependent radical enzyme. Identification of a dynamic interface between the cobalamin and pyridoxal 5'-phosphate-binding domains, overview. Following ligand binding-induced cleavage of the Lys629-pyridoxal 5'-phosphate covalent bond, dynamic motion of the cobalamin-binding domain leads to conformational sampling of the available space. This supports radical catalysis through transient formation of a catalytically competent active state. Crucially, it appears that the formation of the state containing both a substrate/product radical and Co(II) does not restrict cobalamin domain motion | Acetoanaerobium sticklandii | |
pyridoxal 5'-phosphate | - |
Acetoanaerobium sticklandii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.72 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant P343W | Acetoanaerobium sticklandii | |
1.05 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant G339W | Acetoanaerobium sticklandii | |
1.36 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant E338A | Acetoanaerobium sticklandii | |
4.11 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant I424E | Acetoanaerobium sticklandii | |
14.97 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant D627A | Acetoanaerobium sticklandii | |
15.48 | - |
D-ornithine | pH 8.5, 25°C, recombinant beta-subunit mutant C700S | Acetoanaerobium sticklandii | |
15.71 | - |
D-ornithine | pH 8.5, 25°C, recombinant wild-type enzyme | Acetoanaerobium sticklandii |