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Information on EC 1.3.1.118 - meromycolic acid enoyl-[acyl-carrier-protein] reductase and Organism(s) Mycobacterium tuberculosis and UniProt Accession P9WGR1
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1.3.1.118 meromycolic acid enoyl-[acyl-carrier-protein] reductaseIUBMB Comments
InhA is a component of the fatty acid synthase (FAS) II system of Mycobacterium tuberculosis, catalysing an enoyl-[acyl-carrier-protein] reductase step. The enzyme acts on very long and unsaturated fatty acids that form the meromycolic component of mycolic acids. It extends FASI-derived C20 fatty acids to form C60 to C90 mycolic acids. The enzyme, which forms a homotetramer, is the target of the preferred antitubercular drug isoniazid.
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Word Map
- 1.3.1.118
- tuberculosis
- mycobacterium
- isoniazid
- ethionamide
-
antituberculous
-
isonicotinic
-
drug-targeted
- pharmacology
- fas-ii
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antitubercular
The taxonomic range for the selected organisms is: Mycobacterium tuberculosis
The expected taxonomic range for this enzyme is: Mycobacteriaceae
The expected taxonomic range for this enzyme is: Mycobacteriaceae
Reaction Schemes
Synonyms
inha protein, enoyl-acp reductase inha, 2-trans-enoyl-acyl carrier protein reductase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
InhA
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InhA
InhA
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-
-
-
PATHWAY SOURCE
PATHWAYS
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-
SYSTEMATIC NAME
IUBMB Comments
meromycolyl-[acyl-carrier protein]:NAD+ oxidoreductase
InhA is a component of the fatty acid synthase (FAS) II system of Mycobacterium tuberculosis, catalysing an enoyl-[acyl-carrier-protein] reductase step. The enzyme acts on very long and unsaturated fatty acids that form the meromycolic component of mycolic acids. It extends FASI-derived C20 fatty acids to form C60 to C90 mycolic acids. The enzyme, which forms a homotetramer, is the target of the preferred antitubercular drug isoniazid.
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
trans-2-decenoyl-CoA + NADH + H+
decanoyl-CoA + NAD+
-
-
-
?
trans-2-dodecenoyl-CoA + NADH + H+
dodecanoyl-CoA + NAD+
-
-
-
?
trans-2-hexadecenoyl-CoA + NADH + H+
hexadecanoyl-CoA + NAD+
-
-
-
?
trans-2-octenoyl-CoA + NADH + H+
octanoyl-CoA + NAD+
-
-
-
?
trans-2-octenoyl-[acyl-carrier protein] + NADH + H+
octanoyl-[acyl-carrier protein] + NAD+
-
-
-
?
trans-2-decenoyl-CoA + NADH + H+
decanoyl-CoA + NAD+
-
-
-
-
?
trans-2-dodecenoyl-CoA + NADH + H+
dodecanoyl-CoA + NAD+
-
-
-
-
?
trans-2-dodecenoyl-CoA + NADH + H+
dodecanoyl-CoA + NAD+
-
pre-steady state kinetics and equilibrium data of 2-trans-dodecenoyl-CoA substrate binding to InhA. The results indicate both positive homotropic cooperativity upon substrate binding to InhA, and a bimolecular association process followed by a slow isomerization of the enzyme-substrate binary complex
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-
?
additional information
?
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no activity with crotonoyl-CoA. The enzyme preferentially reduces long-chain 2-trans-enoyl-ACP substrates (12-24 carbons). The two substrates bind to InhA via a sequential kinetic mechanism, with the preferred ordered addition of NADH and the enoyl substrate. The chemical mechanism involves stereospecific hydride transfer of the 4S hydrogen of NADH to the C3 position of the 2-trans-enoyl substrate, followed by protonation at C2 of an enzymestabilized enolate intermediate
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-
?
additional information
?
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-
no activity with crotonoyl-CoA. The enzyme preferentially reduces long-chain 2-trans-enoyl-ACP substrates (12-24 carbons). The two substrates bind to InhA via a sequential kinetic mechanism, with the preferred ordered addition of NADH and the enoyl substrate. The chemical mechanism involves stereospecific hydride transfer of the 4S hydrogen of NADH to the C3 position of the 2-trans-enoyl substrate, followed by protonation at C2 of an enzymestabilized enolate intermediate
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-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NADH
although very flexible, in the wild-type enzyme/NADH complex, the NADH molecule keeps its extended conformation firmly bound to the binding site of the enzyme
NADH
while NADH binding to wild-type InhA is hyperbolic, the InhA mutants bind the cofactor with positive cooperativity, suggesting that the mutations permit access to a second conformational state of the protein
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(1H-indol-5-yl)[4-[(4-methylphenyl)(phenyl)methyl]piperazin-1-yl]methanone
0.015 mM, 74% inhibition
(4-(9H-fluoren-9-yl) piperazin-1-yl)-(4-methylbenzyl)-methanone
0.015 mM, 99% inhibition
(4-(9H-fluoren-9-yl)piperazin-1-yl) (2,5-difluorophenyl)methanone
uncompetitive versus NADH, noncompetitive versus trans-2-dodecenoyl-CoA
(4-(9H-fluoren-9-yl)piperazin-1-yl) (2-fluorophenyl)methanone
uncompetitive versus NADH, noncompetitive versus trans-2-dodecenoyl-CoA
(4-(9H-fluoren-9-yl)piperazin-1-yl) (3-fluorophenyl)methanone
uncompetitive versus NADH, competitive versus trans-2-dodecenoyl-CoA
(4-(9H-Fluoren-9-yl)piperazin-1-yl) (3-tolyl)methanone
uncompetitive versus NADH, noncompetitive versus trans-2-dodecenoyl-CoA
(4-(9H-fluoren-9-yl)piperazin-1-yl) (4-chlorophenyl)methanone
uncompetitive versus NADH, noncompetitive versus trans-2-dodecenoyl-CoA
(4-(9H-fluoren-9-yl)piperazin-1-yl) (4-fluorophenyl)methanone
uncompetitive versus NADH, competitive versus trans-2-dodecenoyl-CoA
(4-(9H-fluoren-9-yl)piperazin-1-yl) (4-methoxyphenyl)methanone
uncompetitive versus NADH, noncompetitive versus trans-2-dodecenoyl-CoA
(4-(9H-fluoren-9-yl)piperazin-1-yl) (4-tolyl)methanone
uncompetitive versus NADH, competitive versus trans-2-dodecenoyl-CoA
(4-(9H-fluoren-9-yl)piperazin-1-yl) (phenyl)methanone
uncompetitive versus NADH, noncompetitive versus trans-2-dodecenoyl-CoA
(4-(9H-fluoren-9-yl)piperazin-1-yl)(indolin-5-yl)methanone
i.e. Genz10850
(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-(1H-indole-5-carbonyl)-methanone
0.015 mM, 84% inhibition
(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-(4-methylbenzyl)-methanone
0.015 mM, 83% inhibition
(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-benzyl-methanone
0.015 mM, 81% inhibition
(4-methylphenyl)[4-[(4-methylphenyl)(phenyl)methyl]piperazin-1-yl]methanone
0.015 mM, 77% inhibition
1-cyclohexyl-N-(2,5-dimethylphenyl)-5-oxopyrrolidine-3-carboxamide
-
1-cyclohexyl-N-(3,5-dichlorophenyl)-5-oxopyrrolidine-3-carboxamide
-
1-cyclohexyl-N-(5'-hydroxy-[1,1':4',1''-terphenyl]-2'-yl)-5-oxopyrrolidine-3-carboxamide
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2-(2-chloro-4-fluorophenyl)-N-(4-((3,5-dimethyl-1H-pyrazol-1-yl)-methyl)phenyl)acetamide
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2-(2-chlorophenoxy)-5-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]phenol
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2-(2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(5-nitrothiazol-2-yl)acetamide
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2-(2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(6-nitrobenzo[d]thiazol-2-yl)acetamide
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2-(2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(thiophen-2-ylmethyl)acetamide
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2-(2-methylphenoxy)-5-[[4-(3-methylphenyl)-1H-1,2,3-triazol-1-yl]methyl]phenol
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2-(4-oxoquinazolin-3(4H)-yl)-N-(thiophen-2-ylmethyl)acetamide
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2-(4-[[4-(2-bromoethyl)-1H-1,2,3-triazol-1-yl]methyl]-2-hydroxyphenoxy)benzonitrile
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2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(2-chloro-5-(trifluoromethyl)phenyl)acetamide
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2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(5-nitrothiazol-2-yl)acetamide
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2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(6-nitrobenzo[d]thiazol-2-yl)acetamide
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2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(furan-2-ylmethyl)acetamide
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2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(thiophen-2-ylmethyl)acetamide
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2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)-N-phenylacetamide
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2-(6-chloro-4-oxoquinazolin-3(4H)-yl)-N-(2-chloro-5-(trifluoromethyl)phenyl)acetamide
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2-(6-chloro-4-oxoquinazolin-3(4H)-yl)-N-(5-nitrothiazol-2-yl)acetamide
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2-(6-chloro-4-oxoquinazolin-3(4H)-yl)-N-(6-nitrobenzo[d]thiazol-2-yl)acetamide
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2-(6-chloro-4-oxoquinazolin-3(4H)-yl)-N-(furan-2-ylmethyl)acetamide
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2-(6-chloro-4-oxoquinazolin-3(4H)-yl)-N-(thiophen-2-ylmethyl)acetamide
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2-(ethanesulfonyl)-6-[2-[(E)-(hydroxyimino)methyl]-4-(trifluoromethyl)phenoxy]-2,3,1-benzodiazaborinin-1(2H)-ol
-
2-(ethanesulfonyl)-7-[2-[(Z)-(hydroxyimino)methyl]-4-(trifluoromethyl)phenoxy]-2,3,1-benzodiazaborinin-1(2H)-ol
diazaborine, in vitro bactericidal activity against replicating bacteria active against several drug-resistant clinical isolates. AN12855 binds to and inhibits the substrate-binding site of InhA in a cofactor-independent manner. It shows good drug exposure after i.v. and oral delivery, with 53% oral bioavailability. Delivered orally, AN12855 exhibits dose-dependent efficacy in both an acute and chronic murine model of tuberculosis infection. AN12855 is a promising candidate for the development of new antitubercular agents
2-(o-tolyloxy)-5-hexylphenol
i.e. PT70, slow, tight binding inhibitor. It binds preferentially to the enzyme/NAD+x01complex and has a residence time of 24 min on the target, which is 14000 times longer than that of the rapid reversible inhibitor from which it is derived. The 1.8 A crystal structure of the ternary complex between InhA, NAD+, and PT70 reveals the molecular details of enzyme inhibitor recognition and supports the hypothesis that slow onset inhibition is coupled to ordering of an active site loop, which leads to the closure of the substrate-binding pocket
2-[2-hydroxy-4-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]phenoxy]benzonitrile
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2-[4-[(4-cyclohexyl-1H-1,2,3-triazol-1-yl)methyl]-2-hydroxyphenoxy]benzonitrile
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2-[4-[(4-cyclopentyl-1H-1,2,3-triazol-1-yl)methyl]-2-hydroxyphenoxy]benzonitrile
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2-[4-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]-2-hydroxyphenoxy]benzonitrile
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2-[4-[(4-ethyl-1H-1,2,3-triazol-1-yl)methyl]-2-hydroxyphenoxy]benzonitrile
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4-(trifluoromethyl)-2-(4,5-dihydro-4-(2,4-dinitrophenyl)pyrazol-1-yl)pyrimidine
i.e. Genz8575
4-[[1-hydroxy-2-(methanesulfonyl)-1,2-dihydro-2,3,1-benzodiazaborinin-7-yl]oxy]benzonitrile
-
5-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]-2-(2-methylphenoxy)phenol
-
5-[(4-ethyl-1H-1,2,3-triazol-1-yl)methyl]-2-(2-methylphenoxy)phenol
-
5-[2-[(E)-(hydroxyimino)methyl]phenoxy]-2,1-benzoxaborol-1(3H)-ol
-
5-[[4-(4-chlorophenyl)-1H-1,2,3-triazol-1-yl]methyl]-2-(2-methylphenoxy)phenol
-
7-[2-[(Z)-(hydroxyimino)methyl]-4-(trifluoromethyl)phenoxy]-2-(methanesulfonyl)-2,3,1-benzodiazaborinin-1(2H)-ol
-
N-((4-bromo-1-ethyl-1H-pyrazol-5-yl)methyl)-4-((3,5-dimethyl-1Hpyrazol-1-yl)methyl)benzamide
-
N-(2,3-dichlorophenyl)-4-(1H-pyrrol-1-yl)benzamide
anti-tuberculosis activity
N-(2-aminophenyl)-4-(1H-pyrrol-1-yl)benzamide
anti-tuberculosis activity
N-(2-bromobenzyl)-4-[(3,5-dimethyl-1H-pyrazol-1-yl)methyl]benzamide
-
N-(2-chloro-4-fluorobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
-
N-(2-chloro-4-fluorobenzyl)-4-((4-methylthiazol-2-yl)methyl)-benzamide
-
N-(2-chloro-5-(2-phenylacetamido)benzyl)-4-((3,5-dimethyl-1Hpyrazol-1-yl)methyl)benzamide
-
N-(2-chloro-5-(3-phenylureido)benzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
-
N-(2-chloro-5-(trifluoromethyl)phenyl)-2-(2-methyl-4-oxoquinazolin-3(4H)-yl)acetamide
-
N-(2-chloro-5-(trifluoromethyl)phenyl)-2-(4-oxoquinazolin-3(4H)-yl)acetamide
-
N-(2-chloro-5-aminobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
-
N-(2-chlorobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
-
N-(2-nitrobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
-
N-(2-nitrophenyl)-4-(1H-pyrrol-1-yl)benzamide
anti-tuberculosis activity
N-(2-trifluorobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)-benzamide
-
N-(3-bromobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
-
N-(3-chlorobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
-
N-(3-fluorophenyl)-4-(1H-pyrrol-1-yl)benzamide
anti-tuberculosis activity
N-(3-methanesulfonybenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)-methyl)benzamide
-
N-(3-propan-2-yloxybenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
-
N-(4-chlorophenyl)-4-(1H-pyrrol-1-yl)benzamide
anti-tuberculosis activity
N-(4-fluoro-2-(trifluoromethyl)benzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
-
N-(4-fluorobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
-
N-(5-nitrothiazol-2-yl)-2-(4-oxoquinazolin-3(4H)-yl)acetamide
-
N-(6-nitrobenzo[d]thiazol-2-yl)-2-(4-oxoquinazolin-3(4H)-yl)acetamide
-
N-(benzo[d]thiazol-2-yl)-2-(2-methyl-4-oxoquinazolin-3(4H)-yl)acetamide
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N-(benzo[d]thiazol-2-yl)-2-(4-oxoquinazolin-3(4H)-yl)acetamide
-
N-(benzo[d]thiazol-2-yl)-2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)acetamide
-
N-(benzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
-
N-(furan-2-yl-methyl)-2-(4-oxoquinazolin-3(4H)-yl)acetamide
-
N-(furan-2-ylmethyl)-2-(2-methyl-4-oxoquinazolin-3(4H)-yl)acetamide
-
N-([1,1'-biphenyl]-4-yl)-1-cyclohexyl-5-oxopyrrolidine-3-carboxamide
-
N-[(1-[[3-hydroxy-4-(2-methylphenoxy)phenyl]methyl]-1H-1,2,3-triazol-4-yl)methyl]cyclopropanecarboxamide
-
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(2-methyl-1,3-thiazol-4-yl)methyl]benzamide
-
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(3-methyl-1H-pyrazol-1-yl)methyl]benzamide
-
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(6-methylpyridin-2-yl)-oxy]benzamide
-
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(6-methylpyridin-2-yl)methyl]benzamide
-
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(6-methylpyridin-2-yl)sulfanyl]benzamide
-
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(dimethyl-1,3-thiazol-2-yl)methyl]benzamide
-
N-[(2-chloro-4-fluorophenyl)methyl]-4-[2-(3,5-dimethyl-1H-pyrazol-1-yl)ethyl]benzamide
-
N-[(4-fluorophenyl)methyl]-4-[[2-methyl-5-(2,2,2-trifluoroethyl)furan-3-yl]methyl]benzamide
-
pentacyano(isoniazid)ferrate(II)
the inorganic complex inhibits both wild-type and isoniazid-resistant Ile21Val mutants of InhA and this inactivation did not require activation by KatG. Molecular dynamics simulations show that the interaction of pentacyano(isoniazid)ferrate(II) with InhA leads to macromolecular instabilities reflected in the long time necessary for simulation convergence. These instabilities are mainly due to perturbation of the substrate binding loop, particularly the partial denaturation of helices alpha6 and alpha7
prothionamide
the prodrug requires activation by EthA, a flavin-dependent monooxygenase. According to the crystal structure of InhA with bound prothionamide-NAD adduct, the propyl-isonicotinic-acyl moiety is located in a hydrophobic pocket formed by the rearrangement of the side chain of Phe149, and an aromatic ringstacking interaction with the pyridine ring
Trp-Tyr-Trp
structure-based computer modelling approach to design a tripeptide inhibitor. Docking studies indicate that the designed peptide has potency 100 times higher than the best known inhibitor. The results suggest that the designed inhibitor is a suitable lead compound for the development of novel anti-TB drugs
[4-(9H-fluoren-9-yl) piperazin-1-yl]-benzyl-methanone
0.015 mM, 97% inhibition
[4-(9H-fluoren-9-yl)piperazin-1-yl](1H-indol-5-yl)methanone
0.015 mM, 94% inhibition
[4-[(4-chlorophenyl)(phenyl)methyl]piperazin-1-yl](1H-indol-5-yl)methanone
0.015 mM, 67% inhibition
[4-[(4-chlorophenyl)(phenyl)methyl]piperazin-1-yl](4-methylphenyl)methanone
0.015 mM, 74% inhibition
[4-[(4-fluorophenyl)(phenyl)methyl]piperazin-1-yl](1H-indol-5-yl)methanone
0.015 mM, 81% inhibition
[4-[(4-fluorophenyl)(phenyl)methyl]piperazin-1-yl](phenyl)methanone
0.015 mM, 81% inhibition
1-(2-furoyl)-4-[3-(2-ethylphenoxy)benzyl]piperazine
-
39.5% residual activity at 0.05 mM
-
1-(2-furoyl)-4-[3-(2-methylphenoxy)benzyl]piperazine
-
41.2% residual activity at 0.05 mM
-
1-(2-furoyl)-4-[3-(phenoxy)benzyl]piperazine
-
KES4, potent inhibitor, 68% residual activity at 0.05 mM
-
1-(2-furoyl)-4-[3-[2-(sec-butyl)phenoxy]benzyl]piperazine
-
48.8% residual activity at 0.05 mM
-
1-(2-furoyl)-4-[3-[2-(tert-butyl)phenoxy]benzyl]piperazine
-
66.5% residual activity at 0.05 mM
-
1-(2-furoyl)-4-[3-[3-(tert-butyl)phenoxy]benzyl]piperazine
-
63.9% residual activity at 0.05 mM
-
1-(2-furoyl)-4-[3-[3-(trifluoromethyl)phenoxy]benzyl]piperazine
-
45.4% residual activity at 0.05 mM
-
1-(2-furoyl)-4-[3-[4-(methoxycarbonylmethyl)phenoxy]benzyl]piperazine
-
83.4% residual activity at 0.05 mM
-
isoniazid-coenzyme adduct
-
inhibition by several types of isoniazid-coenzyme adducts coexisting in solution is discussed in relation with the structure of the coenzyme, the stereochemistry of the adducts, and their existence as both open and cyclic forms
-
Ethionamide
the indirect inhibitor forms a covalent adduct with the cofactor
Ethionamide
the prodrug requires activation by EthA, a flavin-dependent monooxygenase. According to the crystal structure of InhA with bound ethionamide-NAD adduct, the ethyl-isonicotinic-acyl moiety is located in a hydrophobic pocket formed by the rearrangement of the side chain of Phe149, and an aromatic ringstacking interaction with the pyridine ring
isoniazid
a pro-drug, which is oxidatively activated in vivo by the katG-encoded mycobacterial catalase peroxidase to generate an isonicotinoyl radical. This highly reactive species then reacts nonenzymatically with the cellular pyridine nucleotide coenzymes, NAD+ and NADP+, to generate 12 isonicotinoyl-NAD(P)+ adducts. Of these, the acyclic 4S isomer of isoniazid-NAD+ and the acyclic 4R isomer of isoniazid-NADP+ inhibit the inhA-encoded enoyl-ACP reductase
isoniazid
as a pro-drug, isoniazid requires activation by KatG, a catalase-peroxidase enzyme with dual activities of catalase and peroxidase oxidizing isoniazid to an acyl radical binding to position 4 of nicotinamide adenine dinucleotide (NAD) to form an active isoniazid-NAD adduct. Addition of the isonicotinoyl radical to position 4 of the nicotinamide ring can result in two stereoisomersi n which only 4(S) isomers of isoniazid-NAD adduct possess potent activity
isoniazid
inhibits InhA via formation of a covalent adduct with NAD+. KatG, the mycobacterial catalase-peroxidase, is essential for isoniazid activation. While cross-linking studies indicate that enzyme inhibition causes dissociation of the InhA tetramer into dimers, analytical ultracentrifugation and size exclusion chromatography reveal that ligand binding causes a conformational change in the protein that prevents cross-linking across one of the dimer-dimer interfaces in the InhA tetramer
isoniazid
inhibits through the formation of an INH-NAD adduct which is a slow-onset inhibitor of InhA
isoniazid
prodrug which is biologically activated by the Mycobacterium tuberculosis catalase-peroxidase KatG enzyme. The activation reaction promotes the formation of an isonicotinyl-NAD adduct which inhibits the InhA enzyme, resulting in reduction of mycolic acid biosynthesis
isoniazid
the indirect inhibitor forms a covalent adduct with the cofactor, leading compound for antitubercular drug therapy
isoniazid
-
acts on the mycobacterial cell wall by preventing the FAS-II system from producing long-chain fatty acid precursors for mycolic acid biosynthesis
isoniazid
-
fast and efficient inhibition of InhA in the presence of NADH and INH using MnIII-pyrophosphate as nonenzymatic reagent. This chemical oxidant might be a useful tool for further mechanistic studies of isoniazid activation in attempts to establish the exact structures of isoniazid reactive species and InhA inhibitor complex(es).
triclosan
-
demonstration of triclosan inhibition of InhA in yeast represents a meaningful variation in studying this effect in mycobacteria, because it occurrs without the potentially confusing aspects of perturbing protein-protein interactions which are presumed vital to mycobacterial FASII, inactivating other important enzymes or eliciting a dedicated transcriptional response in Mycobacterium tuberculosis
additional information
a series of piperazine derivatives is synthesized and screened as MtInhA inhibitors, which results in the identification of compounds with IC50 values in the submicromolar range
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additional information
overview of 80 available crystal structures of wild-type and mutant InhA, in its apo form, in complex with its cofactor, with an analogue of its natural ligands (C16 fatty acid and/or NADH) or with inhibitors
-
additional information
-
overview of 80 available crystal structures of wild-type and mutant InhA, in its apo form, in complex with its cofactor, with an analogue of its natural ligands (C16 fatty acid and/or NADH) or with inhibitors
-
additional information
twenty eight 2-(4-oxoquinazolin-3(4H)-yl)acetamide derivatives are synthesized and evaluated for their in vitro Mycobacterium tuberculosis InhA inhibition. Compounds are evaluated for their in vitro activity against drug sensitive and resistant Mycobacterium tuberculosis strains and cytotoxicity against RAW 264.7 cell line. Compounds are docked at the active site of InhA to understand their binding mode and differential scanning fluorimetry is performed to ascertain their protein interaction and stability
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additional information
-
SAR studies on novel inhibitor scaffolds. Inhibitor scaffolds include the diaryl ethers, pyrrolidine carboxamides, piperazine indoleformamides, pyrazoles, arylamides, fatty acids, and imidazopiperidines, all of which form ternary complexes with InhA and the NAD cofactor, as well as isoniazid and the diazaborines which covalently modify the cofactor. Analysis of the structural data has enabled the development of a common binding mode for the ternary complex inhibitors, which includes a hydrogen bond network, a large hydrophobic pocket and a third size-limited binding area comprised of both polar and non-polar groups. A critical factor in InhA inhibition involves ordering of the substrate binding loop, located close to the active site, and a direct link is proposed between loop ordering and slow onset enzyme inhibition. Slow onset inhibitors have long residence times on the enzyme target, a property that is of critical importance for in vivo activity
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additional information
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the heterologous enzyme is ectopically expressed in a yeast mutant strain from which the native gene encoding the corresponding mitochondrial FASII enzyme is missing.Using an appropriate fungal mitochondrial leader sequence, the mycobacterial protein is directed to the mitochondria, where it can rescue the respiratory growth phenotype of the mutant. The rationale behind the assay is that added antimycolates are foreseen to inhibit the mycobacterial enzyme, thereby recreating the respiratory deficiency of the original mutant, discernible as poor colony formation and growth on glycerol medium
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0015
trans-2-hexadecenoyl-CoA
pH 6.8, 25°C, wild-type enzyme
0.0045
NADH
pH 6.8, 25°C, cosubstrate: trans-2-hexadecenoyl-CoA, wild-type enzyme
0.0058
NADH
pH 6.8, 25°C, cosubstrate: trans-2-dodecenoyl-CoA, wild-type enzyme
0.0076
NADH
pH 6.8, 25°C, cosubstrate: trans-2-octenoyl-CoA, wild-type enzyme
0.0081
NADH
pH 6.8, 25°C, cosubstrate: trans-2-octenoyl-[acyl-carrier protein], wild-type enzyme
0.0373
NADH
pH 6.8, 25°C, cosubstrate: trans-2-octenoyl-[acyl-carrier protein], mutant enzyme S94A
0.065
NADH
pH 6.8, 25°C, cosubstrate: trans-2-octenoyl-CoA, mutant enzyme S94A
0.0196
trans-2-dodecenoyl-CoA
pH 7.5, 25°C, mutant enzyme T266D
0.0203
trans-2-dodecenoyl-CoA
pH 7.5, 25°C, mutant enzyme T266E
0.0409
trans-2-dodecenoyl-CoA
pH 7.5, 25°C, wild-type enzyme
0.048
trans-2-dodecenoyl-CoA
pH 6.8, 25°C, wild-type enzyme
0.0523
trans-2-dodecenoyl-CoA
pH 7.5, 25°C, mutant enzyme T266A
0.688
trans-2-octenoyl-CoA
pH 6.8, 25°C, mutant enzyme S94A
0.002
trans-2-octenoyl-[acyl-carrier protein]
pH 6.8, 25°C, wild-type enzyme
-
0.0027
trans-2-octenoyl-[acyl-carrier protein]
pH 6.8, 25°C, mutant enzyme S94A
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.46
trans-2-dodecenoyl-CoA
pH 7.5, 25°C, mutant enzyme T266D
2.49
trans-2-dodecenoyl-CoA
pH 7.5, 25°C, mutant enzyme T266E
7.12
trans-2-dodecenoyl-CoA
pH 7.5, 25°C, mutant enzyme T266A
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
74.49
trans-2-dodecenoyl-CoA
pH 7.5, 25°C, mutant enzyme T266D
122.7
trans-2-dodecenoyl-CoA
pH 7.5, 25°C, mutant enzyme T266E
130.6
trans-2-dodecenoyl-CoA
pH 7.5, 25°C, wild-type enzyme
136.13
trans-2-dodecenoyl-CoA
pH 7.5, 25°C, mutant enzyme T266A
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.016
2-(2-chlorophenoxy)-5-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]phenol
pH 6.8, 23°C
0.006
2-(2-methylphenoxy)-5-[[4-(3-methylphenyl)-1H-1,2,3-triazol-1-yl]methyl]phenol
pH 6.8, 23°C
0.0071
2-(4-[[4-(2-bromoethyl)-1H-1,2,3-triazol-1-yl]methyl]-2-hydroxyphenoxy)benzonitrile
pH 6.8, 23°C
0.028
2-[2-hydroxy-4-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]phenoxy]benzonitrile
pH 6.8, 23°C
0.0063
2-[4-[(4-cyclohexyl-1H-1,2,3-triazol-1-yl)methyl]-2-hydroxyphenoxy]benzonitrile
pH 6.8, 23°C
0.083
2-[4-[(4-cyclopentyl-1H-1,2,3-triazol-1-yl)methyl]-2-hydroxyphenoxy]benzonitrile
pH 6.8, 23°C
0.099
2-[4-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]-2-hydroxyphenoxy]benzonitrile
pH 6.8, 23°C
0.05
2-[4-[(4-ethyl-1H-1,2,3-triazol-1-yl)methyl]-2-hydroxyphenoxy]benzonitrile
pH 6.8, 23°C
0.0000094
5-hexyl-2-phenoxyphenol
pH and temperature not specified in the publication
0.0000011
5-octyl-2-phenoxyphenol
pH and temperature not specified in the publication
0.0000118
5-pentyl-2-phenoxyphenol
pH and temperature not specified in the publication
0.00003
5-tetradecyl-2-phenoxyphenol
pH and temperature not specified in the publication
0.017
5-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]-2-(2-methylphenoxy)phenol
pH 6.8, 23°C
0.11
5-[(4-ethyl-1H-1,2,3-triazol-1-yl)methyl]-2-(2-methylphenoxy)phenol
pH 6.8, 23°C
0.0019
5-[[4-(4-chlorophenyl)-1H-1,2,3-triazol-1-yl]methyl]-2-(2-methylphenoxy)phenol
pH 6.8, 23°C
0.00013
isoniazid-NADP
pH 7.0, temperature not specified in the publication
1.8
N-[(1-[[3-hydroxy-4-(2-methylphenoxy)phenyl]methyl]-1H-1,2,3-triazol-4-yl)methyl]cyclopropanecarboxamide
pH 6.8, 23°C
0.0002
triclosan
pH and temperature not specified in the publication
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0004
(4-(9H-fluoren-9-yl) piperazin-1-yl)-(4-methylbenzyl)-methanone
Mycobacterium tuberculosis
23°C, pH not specified in the publication
0.00157
(4-(9H-fluoren-9-yl)piperazin-1-yl) (2,5-difluorophenyl)methanone
Mycobacterium tuberculosis
pH 7.0, 25°C
0.000361
(4-(9H-fluoren-9-yl)piperazin-1-yl) (2-fluorophenyl)methanone
Mycobacterium tuberculosis
pH 7.0, 25°C
0.00024
(4-(9H-fluoren-9-yl)piperazin-1-yl) (3-fluorophenyl)methanone
Mycobacterium tuberculosis
pH 7.0, 25°C
0.00025
(4-(9H-Fluoren-9-yl)piperazin-1-yl) (3-tolyl)methanone
Mycobacterium tuberculosis
pH 7.0, 25°C
0.00169
(4-(9H-fluoren-9-yl)piperazin-1-yl) (4-chlorophenyl)methanone
Mycobacterium tuberculosis
pH 7.0, 25°C
0.000397
(4-(9H-fluoren-9-yl)piperazin-1-yl) (4-fluorophenyl)methanone
Mycobacterium tuberculosis
pH 7.0, 25°C
0.0029
(4-(9H-fluoren-9-yl)piperazin-1-yl) (4-methoxyphenyl)methanone
Mycobacterium tuberculosis
pH 7.0, 25°C
0.000222
(4-(9H-fluoren-9-yl)piperazin-1-yl) (4-tolyl)methanone
Mycobacterium tuberculosis
pH 7.0, 25°C
0.000183
(4-(9H-fluoren-9-yl)piperazin-1-yl) (phenyl)methanone
Mycobacterium tuberculosis
pH 7.0, 25°C
0.00104
(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-(1H-indole-5-carbonyl)-methanone
Mycobacterium tuberculosis
23°C, pH not specified in the publication
0.00189
(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-(4-methylbenzyl)-methanone
Mycobacterium tuberculosis
23°C, pH not specified in the publication
0.00204
(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-benzyl-methanone
Mycobacterium tuberculosis
23°C, pH not specified in the publication
0.00516
(4-benzylpiperidin-1-yl)(p-tolyl)methanone
Mycobacterium tuberculosis
pH and temperature not specified in the publication
0.00001005
1-cyclohexyl-N-(2,5-dimethylphenyl)-5-oxopyrrolidine-3-carboxamide
Mycobacterium tuberculosis
pH and temperature not specified in the publication
0.00039
1-cyclohexyl-N-(3,5-dichlorophenyl)-5-oxopyrrolidine-3-carboxamide
Mycobacterium tuberculosis
pH and temperature not specified in the publication
0.00014
1-cyclohexyl-N-(5'-hydroxy-[1,1':4',1''-terphenyl]-2'-yl)-5-oxopyrrolidine-3-carboxamide
Mycobacterium tuberculosis
pH and temperature not specified in the publication
0.00004
2-(2-chloro-4-fluorophenyl)-N-(4-((3,5-dimethyl-1H-pyrazol-1-yl)-methyl)phenyl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00616
2-(2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(6-nitrobenzo[d]thiazol-2-yl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00601
2-(2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(thiophen-2-ylmethyl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00892
2-(2-methyl-4-oxoquinazolin-3(4H)-yl)-N-phenylacetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00716
2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(2-chloro-5-(trifluoromethyl)phenyl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00452
2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(furan-2-ylmethyl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00676
2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)-N-(thiophen-2-ylmethyl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00312
2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)-N-phenylacetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00456
2-(6-chloro-4-oxoquinazolin-3(4H)-yl)-N-(2-chloro-5-(trifluoromethyl)phenyl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00312
2-(6-chloro-4-oxoquinazolin-3(4H)-yl)-N-(furan-2-ylmethyl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00712
2-(6-chloro-4-oxoquinazolin-3(4H)-yl)-N-(thiophen-2-ylmethyl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00778
2-(6-chloro-4-oxoquinazolin-3(4H)-yl)-N-phenylacetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.0023
2-(ethanesulfonyl)-6-[2-[(E)-(hydroxyimino)methyl]-4-(trifluoromethyl)phenoxy]-2,3,1-benzodiazaborinin-1(2H)-ol
Mycobacterium tuberculosis
pH 6.8, 23°C
0.00003
2-(ethanesulfonyl)-7-[2-[(Z)-(hydroxyimino)methyl]-4-(trifluoromethyl)phenoxy]-2,3,1-benzodiazaborinin-1(2H)-ol
Mycobacterium tuberculosis
pH 6.8, 23°C
0.079
4-[[1-hydroxy-2-(methanesulfonyl)-1,2-dihydro-2,3,1-benzodiazaborinin-7-yl]oxy]benzonitrile
Mycobacterium tuberculosis
pH 6.8, 23°C
0.012
5-[2-[(E)-(hydroxyimino)methyl]phenoxy]-2,1-benzoxaborol-1(3H)-ol
Mycobacterium tuberculosis
pH 6.8, 23°C
0.044
6-[4-(trifluoromethyl)phenoxy]-2,1-benzoxaborol-1(3H)-ol
Mycobacterium tuberculosis
pH 6.8, 23°C
0.0004
7-[2-[(Z)-(hydroxyimino)methyl]-4-(trifluoromethyl)phenoxy]-2-(methanesulfonyl)-2,3,1-benzodiazaborinin-1(2H)-ol
Mycobacterium tuberculosis
pH 6.8, 23°C
0.00016
Genz10850
Mycobacterium tuberculosis
pH and temperature not specified in the publication
0.0024
GENZ8575
Mycobacterium tuberculosis
pH and temperature not specified in the publication
0.00009
N-((4-bromo-1-ethyl-1H-pyrazol-5-yl)methyl)-4-((3,5-dimethyl-1Hpyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00054
N-(2-bromobenzyl)-4-[(3,5-dimethyl-1H-pyrazol-1-yl)methyl]benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.0005
N-(2-chloro-4-fluorobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00006
N-(2-chloro-4-fluorobenzyl)-4-((4-methylthiazol-2-yl)methyl)-benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00008
N-(2-chloro-5-(2-phenylacetamido)benzyl)-4-((3,5-dimethyl-1Hpyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00009
N-(2-chloro-5-(3-phenylureido)benzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00906
N-(2-chloro-5-(trifluoromethyl)phenyl)-2-(2-methyl-4-oxoquinazolin-3(4H)-yl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00816
N-(2-chloro-5-(trifluoromethyl)phenyl)-2-(4-oxoquinazolin-3(4H)-yl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00035
N-(2-chloro-5-aminobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00126
N-(2-chlorobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00251
N-(2-nitrobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.001
N-(2-trifluorobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)-benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00158
N-(3-bromobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00158
N-(3-chlorobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00251
N-(3-methanesulfonybenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)-methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00158
N-(3-propan-2-yloxybenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00025
N-(4-fluoro-2-(trifluoromethyl)benzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.0031
N-(4-fluorobenzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00512
N-(6-nitrobenzo[d]thiazol-2-yl)-2-(4-oxoquinazolin-3(4H)-yl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00836
N-(benzo[d]thiazol-2-yl)-2-(6-chloro-2-methyl-4-oxoquinazolin-3(4H)-yl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00593
N-(benzyl)-4-((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00686
N-(furan-2-ylmethyl)-2-(2-methyl-4-oxoquinazolin-3(4H)-yl)acetamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00039
N-([1,1'-biphenyl]-4-yl)-1-cyclohexyl-5-oxopyrrolidine-3-carboxamide
Mycobacterium tuberculosis
pH and temperature not specified in the publication
0.00009 - 0.00325
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(2-methyl-1,3-thiazol-4-yl)methyl]benzamide
0.0005
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(3-methyl-1H-pyrazol-1-yl)methyl]benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.0034
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(6-methylpyridin-2-yl)-oxy]benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00005
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(6-methylpyridin-2-yl)methyl]benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00006
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(6-methylpyridin-2-yl)sulfanyl]benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.0014
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(dimethyl-1,3-thiazol-2-yl)methyl]benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00032
N-[(2-chloro-4-fluorophenyl)methyl]-4-[2-(3,5-dimethyl-1H-pyrazol-1-yl)ethyl]benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00012
N-[(4-fluorophenyl)methyl]-4-[[2-methyl-5-(2,2,2-trifluoroethyl)furan-3-yl]methyl]benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00009
[4-(9H-fluoren-9-yl) piperazin-1-yl]-benzyl-methanone
Mycobacterium tuberculosis
23°C, pH not specified in the publication
0.0002
[4-(9H-fluoren-9-yl)piperazin-1-yl](1H-indol-5-yl)methanone
Mycobacterium tuberculosis
23°C, pH not specified in the publication
0.0034
1-(2-furoyl)-4-[3-(2-ethylphenoxy)benzyl]piperazine
Mycobacterium tuberculosis
-
at pH 6.8 and 25°C
-
0.0078
1-(2-furoyl)-4-[3-(2-methylphenoxy)benzyl]piperazine
Mycobacterium tuberculosis
-
at pH 6.8 and 25°C
-
0.0096
1-(2-furoyl)-4-[3-(phenoxy)benzyl]piperazine
Mycobacterium tuberculosis
-
at pH 6.8 and 25°C
-
0.0022
1-(2-furoyl)-4-[3-[2-(sec-butyl)phenoxy]benzyl]piperazine
Mycobacterium tuberculosis
-
at pH 6.8 and 25°C
-
0.005
1-(2-furoyl)-4-[3-[2-(tert-butyl)phenoxy]benzyl]piperazine
Mycobacterium tuberculosis
-
at pH 6.8 and 25°C
-
0.0018
1-(2-furoyl)-4-[3-[3-(tert-butyl)phenoxy]benzyl]piperazine
Mycobacterium tuberculosis
-
at pH 6.8 and 25°C
-
0.0228
1-(2-furoyl)-4-[3-[3-(trifluoromethyl)phenoxy]benzyl]piperazine
Mycobacterium tuberculosis
-
at pH 6.8 and 25°C
-
26.8
1-(2-furoyl)-4-[3-[4-(methoxycarbonylmethyl)phenoxy]benzyl]piperazine
Mycobacterium tuberculosis
-
at pH 6.8 and 25°C
-
0.0000053
2-(o-tolyloxy)-5-hexylphenol
Mycobacterium tuberculosis
pH 6.8, 23°C, at 10 nM enzyme concentration
0.0000503
2-(o-tolyloxy)-5-hexylphenol
Mycobacterium tuberculosis
pH 6.8, 23°C, at 100 nM enzyme concentration
0.00009
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(2-methyl-1,3-thiazol-4-yl)methyl]benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
0.00025
N-[(2-chloro-4-fluorophenyl)methyl]-4-[(2-methyl-1,3-thiazol-4-yl)methyl]benzamide
Mycobacterium tuberculosis
pH 6.8, 25°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
physiological function
the enzyme is the causative agent of tuberculosis
metabolism
metabolism
the enzyjme is a member of an FAS-II system that prefers longer chain fatty acylsubstrates for the purpose of synthesizing mycolic acids, a major component of mycobacterial cell walls
metabolism
the enzyme catalyzes the NADH-dependent reduction of long-chain trans-2-enoyl-ACP fatty acids, an intermediate in mycolic acid biosynthesis
metabolism
the enzyme is a member of the mycobacterial type II dissociated fatty acid biosynthesis system
metabolism
the enzyme is an essential enzyme of the mycolic acid biosynthetic pathway
metabolism
the enzyme is involved in mycolic acid biosynthesis
metabolism
the enzyme is involved in the biosynthesis of mycolic acids
metabolism
the enzyme is involved in the mycobacterial fatty acid biosynthesis pathway. It is essential for the survival of Mycobacterium tuberculosis
metabolism
the enzyme is involved in the type II fatty acid biosynthesis pathway of Mycobacterium tuberculosis
metabolism
the enzyme is involved in type II fatty acid biosynthetic pathway
metabolism
-
InhA belongs to the FAS-II system and participates in the mycolic acid pathway. The long-chain fatty acids produced by the FAS-II system serve as precursors for the formation of the meromycolic acids, which represent the C40-C60 main chain of mycolic acids
metabolism
-
key enzyme involved in the biosynthesis of long-chain fatty acids and of mycolic acids, specific components of the mycobacterial cell wall
metabolism
-
the enzyme is involved in mycolic acid biosynthesis. It is part of the dissociative type 2 fatty acid synthase (FASII) system
metabolism
-
the enzyme is involved on the biosynthetic pathway for mycolic acids
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
phosphoprotein
InhA is phosphorylated in vitro by multiple Ser/Thr kinases on residue Thr266.. Activity of InhA is controlled via phosphorylation. Thr266 is the unique kinase phosphoacceptor, both in vitro and in vivo. The physiological relevance of Thr266 phosphorylation is demonstrated using inhA phosphoablative (T266A) or phosphomimetic (T266D/E) mutants. Enoyl reductase activity is severely impaired in the mimetic mutants in vitro, as a consequence of a reduced binding affinity to NADH. Introduction of inhA_T266D/E fails to complement growth and mycolic acid defects of an inhA-thermosensitive Mycobacterium smegmatis strain, in a similar manner to what is observed following isoniazid treatment. Phosphorylation of InhA may represent an unusual mechanism that allows Mycobacterium tuberculosis to regulate its mycolic acid content, thus offering a new approach to future anti-tuberculosis drug development
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystallization and structure determination of InhA ternary complexes with PT501, PT504, PT506, PT511, PT512 and PT514
crystallization of InhA with isoniazid-NADP bound and the structure of the complex is solved
hanging drop vapor diffusion technique, formation of the ternary enzyme/NAD+x012-(o-tolyloxy)-5-hexylphenol complex
molecular docking studies are performed on the crystal structure of Mycobacterium tuberculosis enoyl reductase (InhA) complexed with 1-cyclohexyl-N-(3,5-dichlorophenyl)-5-oxopyrrolidine-3-carboxamide
overview of 80 available crystal structures of wild-type and mutant InhA, in its apo form, in complex with its cofactor, with an analogue of its natural ligands (C16 fatty acid and/or NADH) or with inhibitor
the crystal structure of the enzyme in complex with NAD+ and trans-2-hexadecenoyl-(N-acetylcysteamine)-thioester reveals that the substrate binds in a general U-shaped conformation, with the trans double bond positioned directly adjacent to the nicotinamide ring of NAD+. The side chain of Tyr158 directly interacts with the thioester carbonyl oxygen of the C16 fatty acyl substrate and therefore can help stabilize the enolate intermediate, proposed to form during substrate catalysis. Hydrophobic residues, primarily from the substrate binding loop (residues 196-219), engulf the fatty acyl chain portion of the substrate. The substrate binding loop of InhA is longer than that of other enoyl-ACP reductases and creates a deeper substrate binding crevice, consistent with the ability of InhA to recognize longer chain fatty acyl substrates
the crystal structure of the enzyme in complex with the inhibitor N-(4-methylbenzoyl)-4-benzylpiperidine reveals the binding mode of the inhibitor within the active site
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
I16T
I21V
I47T
similar to wild-type InhA, cross-linking of the isoniazid resistant mutant gives three bands on SDS-PAGE assigned to monomer, dimer, and tetrameric forms of the protein. The inhibition of the enzyme with the isoniazid-NAD adduct results in loss of the band assigned to tetramer. In contrast, cross-linking in the presence of saturating concentrations of NADH yields a lower amount of the tetramer upon SDS-PAGE
S94A
T266A
phosphoablative mutant with activity similar to wild-type enzyme
T266D
phosphomimetic mutant with strongly reduced activity (31.4% compared to wild-type enzyme), introduction of inhA_T266D fails to complement growth and mycolic acid defects of an inhA-thermosensitive Mycobacterium smegmatis strain, in a similar manner to what is observed following isoniazid treatment
T266E
phosphomimetic mutant with strongly reduced activity (29.5% compared to wild-type enzyme), introduction of inhA_T266E fails to complement growth and mycolic acid defects of an inhA-thermosensitive Mycobacterium smegmatis strain, in a similar manner to what is observed following isoniazid treatment
Y158A
mutation improves the KM for the cofactor by a factor of 13
Y158F
mutation improves the KM for the cofactor by a factor of 33
I16T
mutation in the glycine-rich loop. Although very flexible, in the wild-type enzyme/NADH complex, the NADH molecule keeps its extended conformation firmly bound to the binding site of the enzyme. In the mutant complex, the NADH pyrophosphate moiety undergoes considerable conformational changes, reducing its interactions with its binding site and probably indicating the initial phase of ligand expulsion from the cavity
I21V
mutation in the glycine-rich loop. Although very flexible, in the wild-type enzyme/NADH complex, the NADH molecule keeps its extended conformation firmly bound to the binding site of the enzyme. In the mutant complex, the NADH pyrophosphate moiety undergoes considerable conformational changes, reducing its interactions with its binding site and probably indicating the initial phase of ligand expulsion from the cavity
I21V
similar to wild-type InhA, cross-linking of the isoniazid resistant mutant gives three bands on SDS-PAGE assigned to monomer, dimer, and tetrameric forms of the protein. The inhibition of the enzyme with the isoniazid-NAD adduct results in loss of the band assigned to tetramer. In contrast, cross-linking in the presence of saturating concentrations of NADH yields a lower amount of the tetramer upon SDS-PAGE
S94A
mutation confers resistance to both isoniazid and ethionamide. Binding of NADH to the mutant is altered
S94A
similar to wild-type InhA, cross-linking of the isoniazid resistant mutant gives three bands on SDS-PAGE assigned to monomer, dimer, and tetrameric forms of the protein. The inhibition of the enzyme with the isoniazid-NAD adduct results in loss of the band assigned to tetramer. In contrast, cross-linking in the presence of saturating concentrations of NADH yields a lower amount of the tetramer upon SDS-PAGE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed with N-terminal hexa-histidine motifs in Escherichia coli BL21 (DE3) pLysS cells
expression of InHA-6xHis protein in Escherichia coli (BL21)
the inhA gene is cloned into the pMK1 mycobacterial expression vector under the control of the strong promoter hsp60. The resulting construct is used to transform Mycobacterium bovis BCG Pasteur in order to allow overproduction of recombinant His-tagged InhA
expression Saccharomyces cerevisiae etr1D cells lacking Etr1p, the 2-trans-enoyl-thioester reductase of mitochondrial type 2 fatty acid synthase. Yeast mitochondria are used as a surrogate compartment for hosting the drug-target protein InhA from mycobacterial FASII. The heterologous enzyme is ectopically expressed in a yeast mutant strain from which the native gene encoding the corresponding mitochondrial FASII enzyme is missing. Using an appropriate fungal mitochondrial leader sequence, the mycobacterial protein is directed to the mitochondria, where it can rescue the respiratory growth phenotype of the mutant. The rationale behind the assay is that added antimycolates are foreseen to inhibit the mycobacterial enzyme, thereby recreating the respiratory deficiency of the original mutant, discernible as poor colony formation and growth on glycerol medium
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pharmacology
pharmacology
-
structural investigations of reactive isoniazid species in order to promote the design of new inhibitors of InhA as potential antituberculous drugs
pharmacology
the enzyme is a target for developing novel anti-tubercular agents
pharmacology
the enzyme is a target for the development of new anti-tubercular drugs
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Nguyen, M.; Quemard, A.; Broussy, S.; Bernadou, J.; Meunier, B.
Mn(III) pyrophosphate as an efficient tool for studying the mode of action of isoniazid on the InhA protein of Mycobacterium tuberculosis
Antimicrob. Agents Chemother.
46
2137-2144
2002
Mycobacterium tuberculosis
Quemard, A.; Sacchettini, J.; Dessen, A.; Vilcheze, C.; Bittman, R.; Jacobs, W.J.; Blanchard, J.
Enzymatic characterization of the target for isoniazid in Mycobacterium tuberculosis
Biochemistry
34
8235-8241
1995
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis, Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Joshi, S.; Dixit, S.; Basha, J.; Kulkarni, V.; Aminabhavi, T.; Nadagouda, M.; Lherbet, C.
Pharmacophore mapping, molecular docking, chemical synthesis of some novel pyrrolyl benzamide derivatives and evaluation of their inhibitory activity against enoyl-ACP reductase (InhA) and Mycobacterium tuberculosis
Bioorg. Chem.
81
440-453
2018
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis H37Rv (P9WGR1)
He, X.; Alian, A.; Ortiz de Montellano, P.
Inhibition of the Mycobacterium tuberculosis enoyl acyl carrier protein reductase InhA by arylamides
Bioorg. Med. Chem.
15
6649-6658
2007
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Schroeder, E.; Basso, L.; Santos, D.; De Souza, O.
Molecular dynamics simulation studies of the wild-type, I21V, and I16T mutants of isoniazid-resistant Mycobacterium tuberculosis enoyl reductase (InhA) in complex with NADH Toward the understanding of NADH-InhA different affinities
Biophys. J.
89
876-884
2005
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Subba Rao, G.; Vijayakrishnan, R.; Kumar, M.
Structure-based design of a novel class of potent inhibitors of InhA, the enoyl acyl carrier protein reductase from Mycobacterium tuberculosis A computer modelling approach
Chem. Biol. Drug Des.
72
444-449
2008
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Nguyen, M.; Quemard, A.; Marrakchi, H.; Bernadou, J.; Meunier, B.
The nonenzymatic activation of isoniazid by MnIII-pyrophosphate in the presence of NADH produces the inhibition of the enoyl-ACP reductase InhA from Mycobacterium tuberculosis
Chemistry
4
35-40
2001
Mycobacterium tuberculosis
-
Guardia, A.; Gulten, G.; Fernandez, R.; Gomez, J.; Wang, F.; Convery, M.; Blanco, D.; Martinez, M.; Perez-Herran, E.; Alonso, M.; Ortega, F.; Rullas, J.; Calvo, D.; Mata, L.; Young, R.; Sacchettini, J.; Mendoza-Losana, A.; Remuinan, M.
N-Benzyl-4-((heteroaryl)methyl)benzamides a new class of direct NADH-dependent 2-trans enoyl-acyl carrier protein reductase (InhA) inhibitors with antitubercular activity
ChemMedChem
11
687-701
2016
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Pan, P.; Tonge, P.
Targeting InhA, the FASII enoyl-ACP reductase SAR studies on novel inhibitor scaffolds
Curr. Top. Med. Chem.
12
672-693
2012
Mycobacterium tuberculosis
Chollet, A.; Maveyraud, L.; Lherbet, C.; Bernardes-Genisson, V.
An overview on crystal structures of InhA protein Apo-form, in complex with its natural ligands and inhibitors
Eur. J. Med. Chem.
146
318-343
2018
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis, Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Pedgaonkar, G.; Sridevi, J.; Jeankumar, V.; Saxena, S.; Devi, P.; Renuka, J.; Yogeeswari, P.; Sriram, D.
Development of 2-(4-oxoquinazolin-3(4H)-yl)acetamide derivatives as novel enoyl-acyl carrier protein reductase (InhA) inhibitors for the treatment of tuberculosis
Eur. J. Med. Chem.
86
613-627
2014
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Rotta, M.; Pissinate, K.; Villela, A.; Back, D.; Timmers, L.; Bachega, J.; De Souza, O.; Santos, D.; Basso, L.; Machado, P.
Piperazine derivatives Synthesis, inhibition of the Mycobacterium tuberculosis enoyl-acyl carrier protein reductase and SAR studies
Eur. J. Med. Chem.
90
436-447
2015
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Argyrou, A.; Vetting, M.; Blanchard, J.
New insight into the mechanism of action of and resistance to isoniazid Interaction of Mycobacterium tuberculosis enoyl-ACP reductase with INH-NADP
J. Am. Chem. Soc.
129
9582-9583
2007
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Spagnuolo, L.; Eltschkner, S.; Yu, W.; Daryaee, F.; Davoodi, S.; Knudson, S.; Allen, E.; Merino, J.; Pschibul, A.; Moree, B.; Thivalapill, N.; Truglio, J.; Salafsky, J.; Slayden, R.; Kisker, C.; Tonge, P.
Evaluating the contribution of transition-state destabilization to changes in the residence time of triazole-based InhA inhibitors
J. Am. Chem. Soc.
139
3417-3429
2017
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Rozwarski, D.; Vilcheze, C.; Sugantino, M.; Bittman, R.; Sacchettini, J.
Crystal structure of the Mycobacterium tuberculosis enoyl-ACP reductase, InhA, in complex with NAD+ and a C16 fatty acyl substrate
J. Biol. Chem.
274
15582-15598
1999
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis, Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Luckner, S.; Liu, N.; Am Ende, C.; Tonge, P.; Kisker, C.
A slow, tight binding inhibitor of InhA, the enoyl-acyl carrier protein reductase from Mycobacterium tuberculosis
J. Biol. Chem.
285
14330-14337
2010
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Vasconcelos, I.; Basso, L.; Santos, D.
Kinetic and equilibrium mechanisms of substrate binding to Mycobacterium tuberculosis enoyl reductase Implications to function-based antitubercular agent design
J. Braz. Chem. Soc.
21
1503-1508
2010
Mycobacterium tuberculosis
-
Da Costa, A.; Pauli, I.; Dorn, M.; Schroeder, E.; Zhan, C.; De Souza, O.
Conformational changes in 2-trans-enoyl-ACP (CoA) reductase (InhA) from M. tuberculosis induced by an inorganic complex A molecular dynamics simulation study
J. Mol. Model.
18
1779-1790
2012
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Gurvitz, A.
Triclosan inhibition of mycobacterial InhA in Saccharomyces cerevisiae Yeast mitochondria as a novel platform for in vivo antimycolate assays
Lett. Appl. Microbiol.
50
399-405
2010
Mycobacterium tuberculosis
Xia, Y.; Zhou, Y.; Carter, D.; McNeil, M.; Choi, W.; Halladay, J.; Berry, P.; Mao, W.; Hernandez, V.; O'Malley, T.; Korkegian, A.; Sunde, B.; Flint, L.; Woolhiser, L.; Scherman, M.; Gruppo, V.; Hastings, C.; Robertson, G.; Ioerger, T.; Sacchettini, J.
Discovery of a cofactor-independent inhibitor of Mycobacterium tuberculosis InhA
Life Sci. Alliance
1
e201800025
2018
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis, Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Marrakchi, H.; Laneelle, G.; Quemard, A.
InhA, a target of the antituberculous drug isoniazid, is involved in a mycobacterial fatty acid elongation system, FAS-II
Microbiology
146
289-296
2000
Mycobacterium tuberculosis, Mycobacterium tuberculosis mc(2)155
Lu, X.; You, Q.; Chen, Y.
Recent progress in the identification and development of InhA direct inhibitors of mycobacterium tuberculosis
Mini Rev. Med. Chem.
10
182-193
2010
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Molle, V.; Gulten, G.; Vilcheze, C.; Veyron-Churlet, R.; Zanella-Cleon, I.; Sacchettini, J.; Jacobs Jr, W.; Kremer, L.
Phosphorylation of InhA inhibits mycolic acid biosynthesis and growth of Mycobacterium tuberculosis
Mol. Microbiol.
78
1591-1605
2010
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Kruh, N.; Rawat, R.; Ruzsicska, B.; Tonge, P.
Probing mechanisms of resistance to the tuberculosis drug isoniazid Conformational changes caused by inhibition of InhA, the enoyl reductase from Mycobacterium tuberculosis
Protein Sci.
16
1617-1627
2007
Mycobacterium tuberculosis (P9WGR1), Mycobacterium tuberculosis ATCC 25618 (P9WGR1)
Jain, S.; Sharma, S.; Sen, D.; Pandya, S.
Enoyl-acyl carrier protein reductase (INHA) A remarkable target to exterminate tuberculosis
Anti-Infect. Agents
19
252-266
2021
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
-
Taira, J.; Umei, T.; Inoue, K.; Kitamura, M.; Berenger, F.; Sacchettini, J.C.; Sakamoto, H.; Aoki, S.
Improvement of the novel inhibitor for Mycobacterium enoyl-acyl carrier protein reductase (InhA) a structure-activity relationship study of KES4 assisted by in silico structure-based drug screening
J. Antibiot.
73
372-381
2020
Mycobacterium tuberculosis
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