Literature summary for 6.3.1.5 extracted from

Arbade, G.K.; Srivastava, S.K.
Cloning, expression, purification, crystallization and preliminary X-ray diffraction studies of NAD synthetase from methicillin-resistant Staphylococcus aureus (2015), Acta Crystallogr. Sect. F, 71, 763-769 .

Application

Application	Comment	Organism
drug development	enzymes involved in NAD metabolism and synthesis are significant drug targets as NAD is a central player in several cellular processes	Staphylococcus aureus

Cloned(Commentary)

Cloned (Comment)	Organism
gene nadE, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)	Staphylococcus aureus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged enzyme enzyme, hanging drop vapour diffusion method, mixing of 0.002-0.004 ml of 10 mg/ml protein in 20 mM TrisHCl, pH 8.5, 200 mM NaCl, and 5% glycerol, with 0.002-0.004 ml of reservoir solution containing 200 mM CaCl2, 100 mM TrisHCl, pH 7.5, 22% PEG 4000, and equilibration against 0.4 ml of reservoir solution, 20°C, 1 week, X-ray diffraction structure determination and analysis at 2.03 A resolution	Staphylococcus aureus

Crystallization (Comment)

Organism

purified recombinant His-tagged enzyme enzyme, hanging drop vapour diffusion method, mixing of 0.002-0.004 ml of 10 mg/ml protein in 20 mM TrisHCl, pH 8.5, 200 mM NaCl, and 5% glycerol, with 0.002-0.004 ml of reservoir solution containing 200 mM CaCl2, 100 mM TrisHCl, pH 7.5, 22% PEG 4000, and equilibration against 0.4 ml of reservoir solution, 20°C, 1 week, X-ray diffraction structure determination and analysis at 2.03 A resolution

Staphylococcus aureus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Staphylococcus aureus

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Staphylococcus aureus	Q5HEK9	-	-
Staphylococcus aureus COL	Q5HEK9	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration	Staphylococcus aureus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + deamido-NAD+ + NH3	-	Staphylococcus aureus	AMP + diphosphate + NAD+	-	?
ATP + deamido-NAD+ + NH3	-	Staphylococcus aureus COL	AMP + diphosphate + NAD+	-	?

Synonyms

Synonyms	Comment	Organism
NAD synthetase	-	Staphylococcus aureus
NadE	-	Staphylococcus aureus
NH3-dependent NAD+ synthetase	-	Staphylococcus aureus

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Staphylococcus aureus

General Information

General Information	Comment	Organism
evolution	the genes encoding the enzymes nicotinic acid mononucleotide adenylyltransferase (NaMNAT), NAD synthetase and NAD kinase responsible for the downstream conversion of nicotinic acid mononucleotide (NaMN) to NAD and NAD(P) are conserved in almost all investigated bacterial genomes	Staphylococcus aureus
metabolism	NAD synthetases are critical as they catalyze the amidation of nicotinic acid adenine dinucleotide (NaAD) to yield the enzyme cofactor NAD, which is the last step in NAD synthesis. NAD synthetase catalyzes the last step in the biosynthesis of nicotinamide adenine dinucleotide, making it a crucial intermediate enzyme linked to the biosynthesis of several amino acids, purine and pyrimidine nucleotides, coenzymes and antibiotics	Staphylococcus aureus