Literature summary for 1.3.1.9 extracted from

Luckner, S.R.; Liu, N.; am Ende, C.W.; Tonge, P.J.; Kisker, C.
A slow, tight binding inhibitor of InhA, the enoyl-acyl carrier protein reductase from Mycobacterium tuberculosis (2010), J. Biol. Chem., 285, 14330-14337.

Application

Application	Comment	Organism
drug development	InhA is an attractive target for the development of drugs against tuberculosis	Mycobacterium tuberculosis

Cloned(Commentary)

Cloned (Comment)	Organism
expression of His-tagged InhA in Escherichia coli BL21(DE3)	Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged InhA in a ternary complex with NAD+ and PT70, 10 mg/ml protein is combined with NAD and PT70 in a molar ratio of 1:5:200, mixing of euqual volumes of protein and reservoir solution, the latter containing 12-16% w/v PEG 4000, 1% DMSO, 100 mM N-(2-acetamido)iminodiacetic acid, pH 6.8, and 100-250 mM ammonium acetate, 22°C, 4 days, X-ray diffraction structure determination and analysis at 1.8-2.1 A resolution, modelling, overview	Mycobacterium tuberculosis

Crystallization (Comment)

Organism

purified recombinant His-tagged InhA in a ternary complex with NAD+ and PT70, 10 mg/ml protein is combined with NAD and PT70 in a molar ratio of 1:5:200, mixing of euqual volumes of protein and reservoir solution, the latter containing 12-16% w/v PEG 4000, 1% DMSO, 100 mM N-(2-acetamido)iminodiacetic acid, pH 6.8, and 100-250 mM ammonium acetate, 22°C, 4 days, X-ray diffraction structure determination and analysis at 1.8-2.1 A resolution, modelling, overview

Mycobacterium tuberculosis

Inhibitors

Inhibitors	Comment	Organism	Structure
2-(o-tolyloxy)-5-hexylphenol	i.e. PT70, a slow, tight binding inhibitor of InhA. PT70 binds preferentially to the InhA-NAD+ complex, binding structure, overview. It has a residence time of 24 min on the target, which is 14000times longer than that of the rapid reversible inhibitor from which it is derived. The slow onset inhibition is coupled to ordering of an active site loop, which leads to the closure of the substrate-binding pocke	Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
an acyl-[acyl-carrier protein] + NAD+	Mycobacterium tuberculosis	-	a trans-2,3-dehydroacyl-[acyl-carrier protein] + NADH + H+	-	?
an acyl-[acyl-carrier protein] + NAD+	Mycobacterium tuberculosis H37Rv	-	a trans-2,3-dehydroacyl-[acyl-carrier protein] + NADH + H+	-	?

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	P9WGR1	-	-
Mycobacterium tuberculosis H37Rv	P9WGR1	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged InhA from Escherichia coli BL21(DE3) by nickel affinity chromatography and gel filtration	Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
an acyl-[acyl-carrier protein] + NAD+	-	Mycobacterium tuberculosis	a trans-2,3-dehydroacyl-[acyl-carrier protein] + NADH + H+	-	?
an acyl-[acyl-carrier protein] + NAD+	-	Mycobacterium tuberculosis H37Rv	a trans-2,3-dehydroacyl-[acyl-carrier protein] + NADH + H+	-	?
additional information	activity detection using trans-2-dodecenyl-CoA and NADH as substrate and cofactor	Mycobacterium tuberculosis	?	-	?
additional information	activity detection using trans-2-dodecenyl-CoA and NADH as substrate and cofactor	Mycobacterium tuberculosis H37Rv	?	-	?

Synonyms

Synonyms	Comment	Organism
enoyl-ACP reductase	-	Mycobacterium tuberculosis
InhA	-	Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.8	-	assay at	Mycobacterium tuberculosis

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	binding structure, overview	Mycobacterium tuberculosis

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
additional information	-	additional information	inhibition kinetics, overview	Mycobacterium tuberculosis
0.000000022	-	2-(o-tolyloxy)-5-hexylphenol	pH 6.8, temperature not specified in the publication	Mycobacterium tuberculosis

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor	Structure
0.0000053	-	pH 6.8, temperature not specified in the publication	Mycobacterium tuberculosis	2-(o-tolyloxy)-5-hexylphenol