title: Structural basis for benzothiazinone-mediated killing of Mycobacterium tuberculosis.
abstract: The benzothiazinone BTZ043 is a tuberculosis drug candidate with nanomolar whole-cell activity. BTZ043 targets the DprE1 catalytic component of the essential enzyme decaprenylphosphoryl-β-D-ribofuranose-2'-epimerase, thus blocking biosynthesis of arabinans, vital components of mycobacterial cell walls. Crystal structures of DprE1, in its native form and in a complex with BTZ043, reveal formation of a semimercaptal adduct between the drug and an active-site cysteine, as well as contacts to a neighboring catalytic lysine residue. Kinetic studies confirm that BTZ043 is a mechanism-based, covalent inhibitor. This explains the exquisite potency of BTZ043, which, when fluorescently labeled, localizes DprE1 at the poles of growing bacteria. Menaquinone can reoxidize the flavin adenine dinucleotide cofactor in DprE1 and may be the natural electron acceptor for this reaction in the mycobacterium. Our structural and kinetic analysis provides both insight into a critical epimerization reaction and a platform for structure-based design of improved inhibitors.
authors.affiliation: Global Health Institute, École Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
authors.forename: João
authors.initials: J
authors.lastname: Neres
authors.forename: Florence
authors.initials: F
authors.lastname: Pojer
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Molteni
authors.forename: Laurent R
authors.initials: LR
authors.lastname: Chiarelli
authors.forename: Neeraj
authors.initials: N
authors.lastname: Dhar
authors.forename: Stefanie
authors.initials: S
authors.lastname: Boy-Röttger
authors.forename: Silvia
authors.initials: S
authors.lastname: Buroni
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Fullam
authors.forename: Giulia
authors.initials: G
authors.lastname: Degiacomi
authors.forename: Anna Paola
authors.initials: AP
authors.lastname: Lucarelli
authors.forename: Randy J
authors.initials: RJ
authors.lastname: Read
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Zanoni
authors.forename: Dale E
authors.initials: DE
authors.lastname: Edmondson
authors.forename: Edda
authors.initials: E
authors.lastname: De Rossi
authors.forename: Maria Rosalia
authors.initials: MR
authors.lastname: Pasca
authors.forename: John D
authors.initials: JD
authors.lastname: McKinney
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Dyson
authors.forename: Giovanna
authors.initials: G
authors.lastname: Riccardi
authors.forename: Andrea
authors.initials: A
authors.lastname: Mattevi
authors.forename: Stewart T
authors.initials: ST
authors.lastname: Cole
authors.forename: Claudia
authors.initials: C
authors.lastname: Binda
mesh_terms: D000995:Antitubercular Agents / Q000737:chemistry* / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry; D018360:Crystallography, X-Ray; D003545:Cysteine / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D005182:Flavin-Adenine Dinucleotide / Q000031:analogs & derivatives / Q000378:metabolism; D005420:Flavoproteins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000378:metabolism; D007700:Kinetics; D008239:Lysine / Q000737:chemistry; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability / Q000187:drug effects*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D020102:Mycobacterium smegmatis / Q000187:drug effects / Q000201:enzymology; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000201:enzymology; D010084:Oxidation-Reduction / Q000187:drug effects; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000737:chemistry; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000187:drug effects; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D013843:Thiazines / Q000737:chemistry* / Q000494:pharmacology*
chemical_list: D000995:Antitubercular Agents; D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D005420:Flavoproteins; D005456:Fluorescent Dyes; D013843:Thiazines; D005182:Flavin-Adenine Dinucleotide; C058805:1,5-dihydro-FAD; D010088:Oxidoreductases; D008239:Lysine; D003545:Cysteine