Literature summary for 2.7.7.18 extracted from

O'Hara, J.K.; Kerwin, L.J.; Cobbold, S.A.; Tai, J.; Bedell, T.A.; Reider, P.J.; Llinas, M.
Targeting NAD+ metabolism in the human malaria parasite Plasmodium falciparum (2014), PLoS ONE, 9, e94061.

Cloned(Commentary)

Cloned (Comment)	Organism
gene PF13_0159, phylogenetic tree, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21 (RIL Codon PLUS), recombinant expression of GFP-tagged enzyme in Plasmodium falciparum ring stage D10 culture cells	Plasmodium falciparum

Protein Variants

Protein Variants	Comment	Organism
D110A	site-directed mutagenesis replacing the conserved catalytic site, inactive mutant	Plasmodium falciparum

Inhibitors

Inhibitors	Comment	Organism
5-(4-bromophenyl)-N'-[(E)-(1-methyl-1H-indol-3-yl)methylidene]-4-oxopentanehydrazide	-	Plasmodium falciparum
5-(4-bromophenyl)-N'-[(E)-(2,3-dimethoxynaphthalen-1-yl)methylidene]-4-oxopentanehydrazide	-	Plasmodium falciparum
5-(4-fluorophenyl)-N'-[(E)-(1-methyl-1H-indol-3-yl)methylidene]-4-oxopentanehydrazide	-	Plasmodium falciparum
5-[(2E)-2-(anthracen-9-ylmethylidene)hydrazinyl]-N-(4-bromophenyl)-5-oxopentanamide	-	Plasmodium falciparum
5-[(2E)-2-(anthracen-9-ylmethylidene)hydrazinyl]-N-(4-fluorophenyl)-5-oxopentanamide	-	Plasmodium falciparum
5-[(2E)-2-(biphenyl-4-ylmethylidene)hydrazinyl]-N-(4-bromophenyl)-5-oxopentanamide	-	Plasmodium falciparum
5-[(2E)-2-(biphenyl-4-ylmethylidene)hydrazinyl]-N-(4-fluorophenyl)-5-oxopentanamide	-	Plasmodium falciparum
5-[(2E)-2-[(2,3-dimethoxynaphthalen-1-yl)methylidene]hydrazinyl]-N-(4-fluorophenyl)-5-oxopentanamide	-	Plasmodium falciparum
5-[(4-bromophenyl)amino]-N'-[(E)-(1-methyl-1H-indol-3-yl)methylidene]hex-5-enehydrazide	-	Plasmodium falciparum
additional information	inhibitor synthesis and in vivo evaluation, MIC50 values determined using a standard SYBR green growth assay on synchronous ring stage parasites, overview	Plasmodium falciparum
N'-[(E)-(2,3-dimethoxynaphthalen-1-yl)methylidene]-5-(4-fluorophenyl)-4-oxopentanehydrazide	-	Plasmodium falciparum
N'-[(E)-anthracen-9-ylmethylidene]-5-(2-bromophenyl)-4-oxopentanehydrazide	-	Plasmodium falciparum
N'-[(E)-anthracen-9-ylmethylidene]-5-(2-hydroxyphenyl)-4-oxopentanehydrazide	-	Plasmodium falciparum
N'-[(E)-anthracen-9-ylmethylidene]-5-(3-bromophenyl)-4-oxopentanehydrazide	-	Plasmodium falciparum
N'-[(E)-anthracen-9-ylmethylidene]-5-(3-hydroxyphenyl)-4-oxopentanehydrazide	-	Plasmodium falciparum
N'-[(E)-anthracen-9-ylmethylidene]-5-(4-bromophenyl)-4-oxopentanehydrazide	-	Plasmodium falciparum
N'-[(E)-anthracen-9-ylmethylidene]-5-(4-fluorophenyl)-4-oxopentanehydrazide	-	Plasmodium falciparum
N'-[(E)-anthracen-9-ylmethylidene]-5-(4-hydroxyphenyl)-4-oxopentanehydrazide	-	Plasmodium falciparum
N'-[(E)-biphenyl-4-ylmethylidene]-5-(4-bromophenyl)-4-oxopentanehydrazide	-	Plasmodium falciparum
N'-[(E)-biphenyl-4-ylmethylidene]-5-(4-fluorophenyl)-4-oxopentanehydrazide	-	Plasmodium falciparum
N-(4-bromophenyl)-5-[(2E)-2-[(2,3-dimethoxynaphthalen-1-yl)methylidene]hydrazinyl]-5-oxopentanamide	-	Plasmodium falciparum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0199	-	ATP	recombinant His6-tagged enzyme, pH 7.5, 37°C	Plasmodium falciparum
0.0358	-	nicotinate beta-D-ribonucleotide	recombinant His6-tagged enzyme, pH 7.5, 37°C	Plasmodium falciparum

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cytoplasm	-	Plasmodium falciparum	5737	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Plasmodium falciparum

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
26000	-	-	Plasmodium falciparum

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + nicotinate beta-D-ribonucleotide	Plasmodium falciparum	-	diphosphate + deamido-NAD+	-	?

Organism

Organism	UniProt	Comment	Textmining
Plasmodium falciparum	Q8IE38	3D7 and D10 clones, gene PF13_0159	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21 (RIL Codon PLUS) by nickel affinity chromatography	Plasmodium falciparum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + nicotinate beta-D-ribonucleotide	-	Plasmodium falciparum	diphosphate + deamido-NAD+	-	?

Subunits

Subunits	Comment	Organism
?	x * 26000, recombinant His6-tagged enzyme, SDS-PAGE	Plasmodium falciparum

Synonyms

Synonyms	Comment	Organism
nicotinate mononucleotide adenylyltransferase	-	Plasmodium falciparum
PfNMNAT	-	Plasmodium falciparum

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Plasmodium falciparum

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Plasmodium falciparum

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Plasmodium falciparum

General Information

General Information	Comment	Organism
evolution	the enzyme is highly diverged from the human homologue, but genetically similar to bacterial NMNATs	Plasmodium falciparum
metabolism	the essential enzyme catalyzes one step in the NAD+ pathway that synthesizes NAD+ from exogenous niacin	Plasmodium falciparum
additional information	residue Asp110 is thought to either hydrogen bond with the adenosine monophosphate during the adenylyltransferase reaction or help with essential Mg+2 coordination during catalysis, the residue has an essential function	Plasmodium falciparum
physiological function	the enzyme is essential for NAD+ metabolism. The enzyme is able to genetically complement the closely related Escherichia coli NMNAT enzyme	Plasmodium falciparum