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Literature summary extracted from

  • Devedjiev, Y.; Symersky, J.; Singh, R.; Jedrzejas, M.; Brouillette, C.; Brouillette, W.; Muccio, D.; Chattopadhyay, D.; DeLucas, L.
    Stabilization of active-site loops in NH3-dependent NAD+ synthetase from Bacillus subtilis (2001), Acta Crystallogr. Sect. D, 57, 806-812.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.1.5 gene OutB, expression in Escherichia coli BL21(DE3) Bacillus subtilis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
6.3.1.5 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

EC Number Metals/Ions Comment Organism Structure
6.3.1.5 Mg2+ required, binding structure and mechanism, 2 binding sites per molecule of enzyme Bacillus subtilis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.1.5 ATP + deamido-NAD+ + NH3 Bacillus subtilis
-
 AMP + diphosphate + NAD+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.1.5 Bacillus subtilis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.1.5 recombinant enzyme from Escherichia coli, over 95% purity Bacillus subtilis

Reaction

EC Number Reaction Comment Organism Reaction ID
6.3.1.5 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
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.1.5 ATP + deamido-NAD+ + NH3
-
 Bacillus subtilis AMP + diphosphate + NAD+
-
 ?
6.3.1.5 ATP + deamido-NAD+ + NH3 dependent on NH3 Bacillus subtilis AMP + diphosphate + NAD+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.3.1.5 NAD synthase
-
 Bacillus subtilis
6.3.1.5 NAD synthetase
-
 Bacillus subtilis
6.3.1.5 NADS
-
 Bacillus subtilis
6.3.1.5 Nicotinamide adenine dinucleotide synthetase
-
 Bacillus subtilis

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.1.5 ATP
-
 Bacillus subtilis