Cloned (Comment) | Organism |
---|---|
gene OutB, expression in Escherichia coli BL21(DE3) | Bacillus subtilis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme, in complex with 4 different substrates: complex I is formed by enzyme and deamido-NAD, complex II is formed by enzyme and ATP, complex III is formed by enzyme, deamido-NAD and ATP, complex is formed by enzyme and substrate analogue alpha,beta-methylene-adenosine triphosphate, crystallization from protein solution: 15 mg/ml protein, 20 mM sodium acetate, pH 5.2, 50 mM MgCl2, 2.5 mM 2-mercaptoethanol, plus equal volume of 0.1 M sodium acetate, pH 5.2, 21-23% PEG 400, 50 mM MgCl2, at room temperature, addition of substrates at different concentrations, formation of complex IV at different pH-values, in the reservoir solution: 50 mM HEPES, pH 7.5, 0.1 M MgCl2, 20-26% PEG 400, 2 mM AMP-CPP, X-ray diffraction structure determination at resolution 1.9-2.3, structure analysis | Bacillus subtilis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required, binding structure and mechanism, 2 binding sites per molecule of enzyme | Bacillus subtilis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + deamido-NAD+ + NH3 | Bacillus subtilis | - |
AMP + diphosphate + NAD+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli, over 95% purity | Bacillus subtilis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + deamido-NAD+ + NH3 = AMP + diphosphate + NAD+ | flexible active site loops of residues 82-7 and 204-225, stabilization of the loop and active site structure, mechanism | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + deamido-NAD+ + NH3 | - |
Bacillus subtilis | AMP + diphosphate + NAD+ | - |
? | |
ATP + deamido-NAD+ + NH3 | dependent on NH3 | Bacillus subtilis | AMP + diphosphate + NAD+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NAD synthase | - |
Bacillus subtilis |
NAD synthetase | - |
Bacillus subtilis |
NADS | - |
Bacillus subtilis |
Nicotinamide adenine dinucleotide synthetase | - |
Bacillus subtilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Bacillus subtilis |