title: Crystal structure of the enoyl-ACP reductase of Mycobacterium tuberculosis (InhA) in the apo-form and in complex with the active metabolite of isoniazid pre-formed by a biomimetic approach.
abstract: InhA is an enoyl-ACP reductase of Mycobacterium tuberculosis implicated in the biosynthesis of mycolic acids, essential constituents of the mycobacterial cell wall. To date, this enzyme is considered as a promising target for the discovery of novel antitubercular drugs. In this work, we describe the first crystal structure of the apo form of the wild-type InhA at 1.80Å resolution as well as the crystal structure of InhA in complex with the synthetic metabolite of the antitubercular drug isoniazid refined to 1.40Å. This metabolite, synthesized in the absence of InhA, is able to displace and replace the cofactor NADH in the enzyme active site. This work provides a unique opportunity to enlighten the structural adaptation of apo-InhA to the binding of the NADH cofactor or of the isoniazid adduct. In addition, a differential scanning fluorimetry study of InhA, in the apo-form as well as in the presence of NAD(+), NADH and INH-NADH was performed showing that binding of the INH-NADH adduct had a strong stabilizing effect.
authors.affiliation: Laboratoire de Chimie de Coordination (LCC), Centre National de la Recherche Scientifique (CNRS), 205 Route de Narbonne, BP 44099, F-31077 Toulouse, Cedex 4, France; Université de Toulouse, Université Paul Sabatier, INPT, F-31077 Toulouse, Cedex 4, France.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Chollet
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale (IPBS), Centre National de la Recherche Scientifique (CNRS), 205 Route de Narbonne, BP 64182, F-31077 Toulouse, France; Université de Toulouse, Université Paul Sabatier, IPBS, Toulouse F-31077, France.
authors.forename: Lionel
authors.initials: L
authors.lastname: Mourey
authors.affiliation: Laboratoire de Synthèse et Physicochimie de Molécules d'Intérêt Biologique (SPCMIB), Centre National de la Recherche Scientifique (CNRS), 118 Route de Narbonne, 31062 Toulouse, Cedex 9, France; Université de Toulouse, Université Paul Sabatier, LSPCMIB, F-31077 Toulouse, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Lherbet
authors.affiliation: Laboratoire de Synthèse et Physicochimie de Molécules d'Intérêt Biologique (SPCMIB), Centre National de la Recherche Scientifique (CNRS), 118 Route de Narbonne, 31062 Toulouse, Cedex 9, France; Université de Toulouse, Université Paul Sabatier, LSPCMIB, F-31077 Toulouse, France.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Delbot
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale (IPBS), Centre National de la Recherche Scientifique (CNRS), 205 Route de Narbonne, BP 64182, F-31077 Toulouse, France; Université de Toulouse, Université Paul Sabatier, IPBS, Toulouse F-31077, France.
authors.forename: Sylviane
authors.initials: S
authors.lastname: Julien
authors.affiliation: Laboratoire de Synthèse et Physicochimie de Molécules d'Intérêt Biologique (SPCMIB), Centre National de la Recherche Scientifique (CNRS), 118 Route de Narbonne, 31062 Toulouse, Cedex 9, France; Université de Toulouse, Université Paul Sabatier, LSPCMIB, F-31077 Toulouse, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Baltas
authors.affiliation: Laboratoire de Chimie de Coordination (LCC), Centre National de la Recherche Scientifique (CNRS), 205 Route de Narbonne, BP 44099, F-31077 Toulouse, Cedex 4, France; Université de Toulouse, Université Paul Sabatier, INPT, F-31077 Toulouse, Cedex 4, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Bernadou
authors.affiliation: Laboratoire de Chimie de Coordination (LCC), Centre National de la Recherche Scientifique (CNRS), 205 Route de Narbonne, BP 44099, F-31077 Toulouse, Cedex 4, France; Université de Toulouse, Université Paul Sabatier, INPT, F-31077 Toulouse, Cedex 4, France.
authors.forename: Geneviève
authors.initials: G
authors.lastname: Pratviel
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale (IPBS), Centre National de la Recherche Scientifique (CNRS), 205 Route de Narbonne, BP 64182, F-31077 Toulouse, France; Université de Toulouse, Université Paul Sabatier, IPBS, Toulouse F-31077, France. Electronic address: laurent.maveyraud@ipbs.fr.
authors.forename: Laurent
authors.initials: L
authors.lastname: Maveyraud
authors.affiliation: Laboratoire de Chimie de Coordination (LCC), Centre National de la Recherche Scientifique (CNRS), 205 Route de Narbonne, BP 44099, F-31077 Toulouse, Cedex 4, France; Université de Toulouse, Université Paul Sabatier, INPT, F-31077 Toulouse, Cedex 4, France. Electronic address: vania.bernardes-genisson@lcc-toulouse.fr.
authors.forename: Vania
authors.initials: V
authors.lastname: Bernardes-Génisson
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D032701:Biomimetics / Q000379:methods; D020134:Catalytic Domain; D007538:Isoniazid / Q000737:chemistry*; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D009243:NAD / Q000737:chemistry; D010088:Oxidoreductases / Q000737:chemistry*; D011485:Protein Binding / Q000502:physiology
chemical_list: D001426:Bacterial Proteins; D009243:NAD; D010088:Oxidoreductases; C085257:InhA protein, Mycobacterium; D007538:Isoniazid
keywords: Apo InhA; Differential scanning fluorimetry; Enoyl-ACP reductase; INH-NADH adduct; Isoniazid; Mycobacterium tuberculosis; X-ray structure