| IED ID | IndEnz0018000616 |
| Enzyme Type ID | peroxidase000616 |
| Protein Name |
Enoyl- acyl-carrier-protein reductase NADH ENR Enoyl-ACP reductase EC 1.3.1.9 FAS-II enoyl-ACP reductase NADH-dependent 2-trans-enoyl-ACP reductase |
| Gene Name | inhA Rv1484 MTCY277.05 |
| Organism | Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv) |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Actinobacteria Actinomycetia (high G+C Gram-positive bacteria) Corynebacteriales Mycobacteriaceae Mycobacterium Mycobacterium tuberculosis complex Mycobacterium tuberculosis Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv) |
| Enzyme Sequence | MTGLLDGKRILVSGIITDSSIAFHIARVAQEQGAQLVLTGFDRLRLIQRITDRLPAKAPLLELDVQNEEHLASLAGRVTEAIGAGNKLDGVVHSIGFMPQTGMGINPFFDAPYADVSKGIHISAYSYASMAKALLPIMNPGGSIVGMDFDPSRAMPAYNWMTVAKSALESVNRFVAREAGKYGVRSNLVAAGPIRTLAMSAIVGGALGEEAGAQIQLLEEGWDQRAPIGWNMKDATPVAKTVCALLSDWLPATTGDIIYADGGAHTQLL |
| Enzyme Length | 269 |
| Uniprot Accession Number | P9WGR1 |
| Absorption | |
| Active Site | |
| Activity Regulation | ACTIVITY REGULATION: InhA activity is controlled via phosphorylation: phosphorylation on Thr-266 decreases InhA activity (5-fold reduction) and likely negatively regulates biosynthesis of mycolic acids and growth of the bacterium (PubMed:20864541, PubMed:21143326). The antitubercular pro-drug isoniazid (INH) is oxidatively activated by the catalase-peroxidase KatG and then covalently binds NAD to form an adduct that inhibits the activity of InhA (Ref.5, PubMed:14623976, PubMed:9417034). The inhibitory adduct is the isonicotinic-acyl-NADH where the isonicotinic-acyl group replaces the 4S (and not the 4R) hydrogen of NADH (PubMed:9417034). Similarly, the antitubercular pro-drugs ethionamide (ETH) and prothionamide (PTH) are activated by the flavoprotein monooxygenase EthA, and forms an adduct with NAD (ETH-NAD and PTH-NAD, respectively) that is a tight-binding inhibitor of InhA (PubMed:17227913). Is inhibited by triclosan and derivatives, pyrazole derivative Genz-8575, indole-5-amide Genz-10850, alkyl diphenyl ethers, pyrrolidine carboxamides, arylamides, pyridomycin, methyl-thiazoles, 4-hydroxy-2-pyridones, and N-benzyl-4-((heteroaryl)methyl)benzamides (PubMed:12606558, PubMed:17163639, PubMed:17034137, PubMed:17723305, PubMed:19130456, PubMed:20200152, PubMed:22987724, PubMed:24107081, PubMed:24616444, PubMed:25568071). Pyridomycin shows a unique mode of InhA inhibition by simultaneously blocking parts of the NADH and the lipid substrate-binding pocket of InhA (PubMed:24292073). Is also inhibited by thiadiazole compounds, that have very attractive antitubercular properties (PubMed:27428438). {ECO:0000269|PubMed:12606558, ECO:0000269|PubMed:14623976, ECO:0000269|PubMed:17034137, ECO:0000269|PubMed:17163639, ECO:0000269|PubMed:17227913, ECO:0000269|PubMed:17723305, ECO:0000269|PubMed:19130456, ECO:0000269|PubMed:20200152, ECO:0000269|PubMed:20864541, ECO:0000269|PubMed:22987724, ECO:0000269|PubMed:24107081, ECO:0000269|PubMed:24292073, ECO:0000269|PubMed:24616444, ECO:0000269|PubMed:25568071, ECO:0000269|PubMed:26934341, ECO:0000269|PubMed:27428438, ECO:0000269|PubMed:9417034, ECO:0000269|Ref.5, ECO:0000305|PubMed:21143326}. |
| Binding Site | BINDING 158; /note="Substrate"; /evidence="ECO:0000269|PubMed:10336454"; BINDING 165; /note="NAD"; /evidence="ECO:0000269|PubMed:10336454, ECO:0000269|PubMed:16647717, ECO:0000269|PubMed:7886450, ECO:0007744|PDB:1BVR, ECO:0007744|PDB:1ENY, ECO:0007744|PDB:2AQ8"; BINDING 194; /note="NAD; via amide nitrogen and carbonyl oxygen"; /evidence="ECO:0000269|PubMed:10336454, ECO:0000269|PubMed:16647717, ECO:0000269|PubMed:7886450, ECO:0007744|PDB:1BVR, ECO:0007744|PDB:1ENY, ECO:0007744|PDB:2AQ8" |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=a 2,3-saturated acyl-[ACP] + NAD(+) = a (2E)-enoyl-[ACP] + H(+) + NADH; Xref=Rhea:RHEA:10240, Rhea:RHEA-COMP:9925, Rhea:RHEA-COMP:9926, ChEBI:CHEBI:15378, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945, ChEBI:CHEBI:78784, ChEBI:CHEBI:78785; EC=1.3.1.9; Evidence={ECO:0000269|PubMed:7599116};PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:10242; Evidence={ECO:0000305|PubMed:7599116}; CATALYTIC ACTIVITY: Reaction=a 2,3-saturated acyl-CoA + NAD(+) = a (2E)-enoyl-CoA + H(+) + NADH; Xref=Rhea:RHEA:18177, ChEBI:CHEBI:15378, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945, ChEBI:CHEBI:58856, ChEBI:CHEBI:65111; Evidence={ECO:0000269|PubMed:10521269, ECO:0000269|PubMed:20864541, ECO:0000269|PubMed:21143326, ECO:0000269|PubMed:22987724, ECO:0000269|PubMed:7599116};PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:18179; Evidence={ECO:0000305|PubMed:7599116}; CATALYTIC ACTIVITY: Reaction=(2E)-octenoyl-[ACP] + H(+) + NADH = NAD(+) + octanoyl-[ACP]; Xref=Rhea:RHEA:41528, Rhea:RHEA-COMP:9635, Rhea:RHEA-COMP:9636, ChEBI:CHEBI:15378, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945, ChEBI:CHEBI:78462, ChEBI:CHEBI:78463; Evidence={ECO:0000269|PubMed:7599116};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:41529; Evidence={ECO:0000305|PubMed:7599116}; CATALYTIC ACTIVITY: Reaction=(2E)-octenoyl-CoA + H(+) + NADH = NAD(+) + octanoyl-CoA; Xref=Rhea:RHEA:63232, ChEBI:CHEBI:15378, ChEBI:CHEBI:57386, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945, ChEBI:CHEBI:62242; Evidence={ECO:0000269|PubMed:20864541, ECO:0000269|PubMed:22987724, ECO:0000269|PubMed:7599116};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:63233; Evidence={ECO:0000305|PubMed:7599116}; CATALYTIC ACTIVITY: Reaction=(2E)-dodecenoyl-CoA + H(+) + NADH = dodecanoyl-CoA + NAD(+); Xref=Rhea:RHEA:45408, ChEBI:CHEBI:15378, ChEBI:CHEBI:57330, ChEBI:CHEBI:57375, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945; Evidence={ECO:0000269|PubMed:10521269, ECO:0000269|PubMed:21143326, ECO:0000269|PubMed:7599116};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:45409; Evidence={ECO:0000305|PubMed:7599116}; CATALYTIC ACTIVITY: Reaction=(2E)-hexadecenoyl-CoA + H(+) + NADH = hexadecanoyl-CoA + NAD(+); Xref=Rhea:RHEA:46072, ChEBI:CHEBI:15378, ChEBI:CHEBI:57379, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945, ChEBI:CHEBI:61526; Evidence={ECO:0000269|PubMed:7599116};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:46073; Evidence={ECO:0000305|PubMed:7599116}; CATALYTIC ACTIVITY: Reaction=(2E)-eicosenoyl-CoA + H(+) + NADH = eicosanoyl-CoA + NAD(+); Xref=Rhea:RHEA:46076, ChEBI:CHEBI:15378, ChEBI:CHEBI:57380, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945, ChEBI:CHEBI:74691; Evidence={ECO:0000269|PubMed:7599116};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:46077; Evidence={ECO:0000305|PubMed:7599116}; CATALYTIC ACTIVITY: Reaction=(2E)-tetracosenoyl-CoA + H(+) + NADH = NAD(+) + tetracosanoyl-CoA; Xref=Rhea:RHEA:46080, ChEBI:CHEBI:15378, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945, ChEBI:CHEBI:65052, ChEBI:CHEBI:74693; Evidence={ECO:0000269|PubMed:7599116};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:46081; Evidence={ECO:0000305|PubMed:7599116}; |
| DNA Binding | |
| EC Number | 1.3.1.9 |
| Enzyme Function | FUNCTION: Enoyl-ACP reductase of the type II fatty acid syntase (FAS-II) system, which is involved in the biosynthesis of mycolic acids, a major component of mycobacterial cell walls (PubMed:25227413). Catalyzes the NADH-dependent reduction of the double bond of 2-trans-enoyl-[acyl-carrier protein], an essential step in the fatty acid elongation cycle of the FAS-II pathway (PubMed:7599116). Shows preference for long-chain fatty acyl thioester substrates (>C16), and can also use 2-trans-enoyl-CoAs as alternative substrates (PubMed:7599116). The mycobacterial FAS-II system utilizes the products of the FAS-I system as primers to extend fatty acyl chain lengths up to C56, forming the meromycolate chain that serves as the precursor for final mycolic acids (PubMed:25227413). {ECO:0000269|PubMed:7599116, ECO:0000303|PubMed:25227413}.; FUNCTION: Is the primary target of the first-line antitubercular drug isoniazid (INH) and of the second-line drug ethionamide (ETH) (PubMed:8284673, PubMed:12406221, PubMed:16906155, PubMed:17227913). Overexpressed inhA confers INH and ETH resistance to M.tuberculosis (PubMed:12406221). The mechanism of isoniazid action against InhA is covalent attachment of the activated form of the drug to the nicotinamide ring of NAD and binding of the INH-NAD adduct to the active site of InhA (PubMed:9417034, PubMed:16906155). Similarly, the ETH-NAD adduct binds InhA (PubMed:17227913). {ECO:0000269|PubMed:12406221, ECO:0000269|PubMed:16906155, ECO:0000269|PubMed:17227913, ECO:0000269|PubMed:9417034, ECO:0000305|PubMed:8284673}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | PATHWAY: Lipid metabolism; mycolic acid biosynthesis. {ECO:0000303|PubMed:25227413}. |
| nucleotide Binding | NP_BIND 20..21; /note="NAD"; /evidence="ECO:0000269|PubMed:10336454, ECO:0000269|PubMed:16647717, ECO:0000269|PubMed:7886450, ECO:0007744|PDB:1BVR, ECO:0007744|PDB:1ENY, ECO:0007744|PDB:2AQ8"; NP_BIND 64..65; /note="NAD"; /evidence="ECO:0000269|PubMed:10336454, ECO:0000269|PubMed:16647717, ECO:0000269|PubMed:7886450, ECO:0007744|PDB:1BVR, ECO:0007744|PDB:1ENY, ECO:0007744|PDB:2AQ8"; NP_BIND 95..96; /note="NAD"; /evidence="ECO:0000269|PubMed:10336454, ECO:0000269|PubMed:16647717, ECO:0000269|PubMed:7886450, ECO:0007744|PDB:1BVR, ECO:0007744|PDB:1ENY, ECO:0007744|PDB:2AQ8" |
| Features | Beta strand (12); Binding site (3); Chain (1); Helix (13); Modified residue (1); Mutagenesis (9); Nucleotide binding (3); Site (2); Turn (4) |
| Keywords | 3D-structure;Antibiotic resistance;Fatty acid biosynthesis;Fatty acid metabolism;Lipid biosynthesis;Lipid metabolism;NAD;Oxidoreductase;Phosphoprotein;Reference proteome |
| Interact With | |
| Induction | |
| Subcellular Location | |
| Modified Residue | MOD_RES 266; /note="Phosphothreonine"; /evidence="ECO:0000269|PubMed:20864541, ECO:0000269|PubMed:21143326" |
| Post Translational Modification | PTM: Is phosphorylated on Thr-266 in vivo. In vitro, can be phosphorylated by multiple Ser/Thr protein kinases (STPK) such as PknA, PknB, PknE, PknH and PknL. Phosphorylation decreases enzymatic activity. {ECO:0000269|PubMed:20864541, ECO:0000269|PubMed:21143326}. |
| Signal Peptide | |
| Structure 3D | X-ray crystallography (102) |
| Cross Reference PDB | 1BVR; 1ENY; 1ENZ; 1P44; 1P45; 1ZID; 2AQ8; 2AQH; 2AQI; 2AQK; 2B35; 2B36; 2B37; 2H9I; 2IDZ; 2IE0; 2IEB; 2IED; 2NSD; 2NTJ; 2NV6; 2PR2; 2X22; 2X23; 3FNE; 3FNF; 3FNG; 3FNH; 3OEW; 3OEY; 3OF2; 4BGE; 4BGI; 4BII; 4BQP; 4BQR; 4COD; 4D0R; 4D0S; 4DQU; 4DRE; 4DTI; 4OHU; 4OIM; 4OXK; 4OXN; 4OXY; 4OYR; 4QXM; 4R9R; 4R9S; 4TRJ; 4TRM; 4TRN; 4TRO; 4TZK; 4TZT; 4U0J; 4U0K; 4UVD; 4UVE; 4UVG; 4UVH; 4UVI; 5COQ; 5CP8; 5CPB; 5CPF; 5G0S; 5G0T; 5G0U; 5G0V; 5G0W; 5JFO; 5MTP; 5MTQ; 5MTR; 5OIF; 5OIL; 5OIM; 5OIN; 5OIT; 5UGS; 5UGT; 5UGU; 6EP8; 6GGM; 6GH1; 6GH4; 6GHN; 6R9W; 6SQ5; 6SQ7; 6SQ9; 6SQB; 6SQD; 6SQL; 6ZKW; 6ZKX; 6ZKY; 6ZKZ; 7E48; |
| Mapped Pubmed ID | 24450589; 27864515; 28151657; 29399991; 30087334; 30217823; 31820972; 32240584; 34677871; 35108401; |
| Motif | |
| Gene Encoded By | |
| Mass | 28,528 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=2.0 uM for 2-trans-octenoyl-ACP (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:7599116}; KM=8.1 uM for NADH (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:7599116}; KM=66 uM for NADH (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:10521269}; KM=19.1 uM for NADH (at pH 6.8) {ECO:0000269|PubMed:20864541}; KM=13.5 uM for NADH (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:22987724}; KM=467 uM for 2-trans-octenoyl-CoA (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:7599116}; KM=528 uM for 2-trans-octenoyl-CoA (at pH 6.8) {ECO:0000269|PubMed:20864541}; KM=48 uM for 2-trans-dodecenoyl-CoA (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:7599116}; KM=27 uM for 2-trans-dodecenoyl-CoA (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:10521269}; KM=40.9 uM for 2-trans-dodecenoyl-CoA (at pH 7.5 and 25 degrees Celsius) {ECO:0000269|PubMed:21143326}; KM=1.5 uM for 2-trans-hexadecenoyl-CoA (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:7599116}; Vmax=2.2 umol/min/mg enzyme for the reduction of 2-trans-octenoyl-ACP (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:7599116}; Vmax=3.6 umol/min/mg enzyme for the reduction of 2-trans-octenoyl-CoA (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:7599116}; Vmax=0.52 umol/min/mg enzyme for the reduction of 2-trans-octenoyl-CoA (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:22987724}; Vmax=15.3 umol/min/mg enzyme for the reduction of 2-trans-octenoyl-CoA (at pH 6.8) {ECO:0000269|PubMed:20864541}; Vmax=5.8 umol/min/mg enzyme for the reduction of 2-trans-dodecenoyl-CoA (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:7599116}; Vmax=11.4 umol/min/mg enzyme for the reduction of 2-trans-dodecenoyl-CoA (at pH 7.5 and 25 degrees Celsius) {ECO:0000269|PubMed:21143326}; Vmax=4.5 umol/min/mg enzyme for the reduction of 2-trans-hexadecenoyl-CoA (at pH 6.8 and 25 degrees Celsius) {ECO:0000269|PubMed:7599116}; Note=kcat is 320.4 min(-1) for the reduction of 2-trans-dodecenoyl-CoA (at pH 7.5 and 25 degrees Celsius) (PubMed:21143326). kcat is 278 min(-1) for the reduction of 2-trans-dodecenoyl-CoA (at pH 6.8 and 25 degrees Celsius) (PubMed:10521269). {ECO:0000269|PubMed:10521269, ECO:0000269|PubMed:21143326}; |
| Metal Binding | |
| Rhea ID | RHEA:10240; RHEA:10242; RHEA:18177; RHEA:18179; RHEA:41528; RHEA:41529; RHEA:63232; RHEA:63233; RHEA:45408; RHEA:45409; RHEA:46072; RHEA:46073; RHEA:46076; RHEA:46077; RHEA:46080; RHEA:46081 |
| Cross Reference Brenda | 1.3.1.118;1.3.1.9; |