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Literature summary for 6.3.1.5 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)

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

Crystallization (Commentary)

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

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Bacillus subtilis
-
-
-

Purification (Commentary)

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

Reaction

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 and Products (Substrate)

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

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

Cofactor

Cofactor Comment Organism Structure
ATP
-
Bacillus subtilis