Literature summary for 6.3.5.1 extracted from

Chuenchor, W.; Doukov, T.I.; Resto, M.; Chang, A.; Gerratana, B.
Regulation of the intersubunit ammonia tunnel in Mycobacterium tuberculosis glutamine-dependent NAD+ synthetase (2012), Biochem. J., 443, 417-426.

Activating Compound

Activating Compound	Comment	Organism	Structure
additional information	the formation of the synthetase intermediate analogues complex, NaAD-AMP intermediate and diphosphatei, triggers structural changes leading to enzyme activation	Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment)	Organism
wild-type mtuNadE in complex with 6-diazo-5-oxo-L-norleucine/NaAD+ and mutant enzymes, hanging drop vapour diffusion method, different combinations of the ligands 3 mM NaAD+, 4 mM ATP, 7 mM adenosine 5'-[alpha,beta-methylene]triphosphate, 20 mM L-glutamine, 6 mM NAD+, 10 mM AMP, 4 mM diphosphate, 100 mM L-glutamate, and 10 mM or 120 mM MgCl2 are used in the co-crystallization experiments, crystallization solution contains 1.2-1.6 M ammonium citrate tribasic, pH 6.5-8.0, with 5-15% v/v glycerol, 20°C, X-ray diffraction structure determination analysis at 2.0-2.85 A resolution, molecular replacement	Mycobacterium tuberculosis

Crystallization (Comment)

Organism

wild-type mtuNadE in complex with 6-diazo-5-oxo-L-norleucine/NaAD+ and mutant enzymes, hanging drop vapour diffusion method, different combinations of the ligands 3 mM NaAD+, 4 mM ATP, 7 mM adenosine 5'-[alpha,beta-methylene]triphosphate, 20 mM L-glutamine, 6 mM NAD+, 10 mM AMP, 4 mM diphosphate, 100 mM L-glutamate, and 10 mM or 120 mM MgCl2 are used in the co-crystallization experiments, crystallization solution contains 1.2-1.6 M ammonium citrate tribasic, pH 6.5-8.0, with 5-15% v/v glycerol, 20°C, X-ray diffraction structure determination analysis at 2.0-2.85 A resolution, molecular replacement

Mycobacterium tuberculosis

Protein Variants

Protein Variants	Comment	Organism
C176A	site-directed mutagenesis, inactive mutant, analysis of ligand binding structures	Mycobacterium tuberculosis
L486A	site-directed mutagenesis	Mycobacterium tuberculosis
L486F	site-directed mutagenesis	Mycobacterium tuberculosis
Y58A	site-directed mutagenesis	Mycobacterium tuberculosis

Inhibitors

Inhibitors	Comment	Organism	Structure
adenosine 5'-[alpha,beta-methylene]triphosphate	i.e. AMP-CPP, nonhydrolysable analogue of ATP	Mycobacterium tuberculosis
nicotinic acid-adenine dinucleotide	i.e. NaAD+	Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.1	2	ATP	pH and temperature not specified in the publication	Mycobacterium tuberculosis
1.5	-	L-glutamine	pH and temperature not specified in the publication	Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + deamido-NAD+ + L-glutamine + H2O	Mycobacterium tuberculosis	-	AMP + diphosphate + NAD+ + L-glutamate	-	?
ATP + deamido-NAD+ + L-glutamine + H2O	Mycobacterium tuberculosis H37Rv	-	AMP + diphosphate + NAD+ + L-glutamate	-	?

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	P9WJJ3	gene nadE	-
Mycobacterium tuberculosis H37Rv	P9WJJ3	gene nadE	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + deamido-NAD+ + L-glutamine + H2O	-	Mycobacterium tuberculosis	AMP + diphosphate + NAD+ + L-glutamate	-	?
ATP + deamido-NAD+ + L-glutamine + H2O	-	Mycobacterium tuberculosis H37Rv	AMP + diphosphate + NAD+ + L-glutamate	-	?

Synonyms

Synonyms	Comment	Organism
glutamine-dependent NAD+ synthetase	-	Mycobacterium tuberculosis
mtuNadE	-	Mycobacterium tuberculosis
NAD+ synthetase	-	Mycobacterium tuberculosis

Cofactor

Cofactor	Comment	Organism	Structure
ATP	structure of the ATP-binding site at the mtuNadE synthetase domain, overview	Mycobacterium tuberculosis

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.7	-	adenosine 5'-[alpha,beta-methylene]triphosphate	pH and temperature not specified in the publication	Mycobacterium tuberculosis

General Information

General Information	Comment	Organism
evolution	most residues lining the ATP-binding site are conserved among the glutamine-dependent NAD+ synthetases, whereas residues not conserved, such as Leu399 andGly366, interact with the adenine ring and the adenylyl ribose with the backbone oxygen and nitrogen	Mycobacterium tuberculosis
additional information	the enzyme contains glutaminase and synthetase active sites, structures and ligand binding, overview. The ATP-binding site is located in a deep cleft formed solely by a single subunit next to the nicotinic acidadenine dinucleotide-binding site	Mycobacterium tuberculosis