title: Structure and Substrate Recognition of the Bottromycin Maturation Enzyme BotP.
abstract: The bottromycins are a family of highly modified peptide natural products, which display potent antimicrobial activity against Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus. Bottromycins have recently been shown to be ribosomally synthesized and post-translationally modified peptides (RiPPs). Unique amongst RiPPs, the precursor peptide BotA contains a C-terminal "follower" sequence, rather than the canonical N-terminal "leader" sequence. We report herein the structural and biochemical characterization of BotP, a leucyl-aminopeptidase-like enzyme from the bottromycin pathway. We demonstrate that BotP is responsible for the removal of the N-terminal methionine from the precursor peptide. Determining the crystal structures of both apo BotP and BotP in complex with Mn2+allowed us to model a BotP/substrate complex and to rationalize substrate recognition. Our data represent the first step towards targeted compound modification to unlock the full antibiotic potential of bottro- mycin.
authors.affiliation: School of Chemistry and Biomedical Sciences Research Centre, University of St. Andrews, North Haugh, St. Andrews, KY16 9ST, UK.
authors.forename: Greg
authors.initials: G
authors.lastname: Mann
authors.affiliation: Roger Adams Laboratory 156, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, IL, 61801, USA.
authors.forename: Liujie
authors.initials: L
authors.lastname: Huo
authors.affiliation: Workgroup Structural Biology of Biosynthetic Enzymes, Helmholtz Institute for Pharmaceutical Research, Helmholtz Centre for Infection Research, Saarland University, Universitätscampus E8 1, 66123, Saarbrücken, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Adam
authors.affiliation: School of Chemistry and Biomedical Sciences Research Centre, University of St. Andrews, North Haugh, St. Andrews, KY16 9ST, UK.
authors.forename: Brunello
authors.initials: B
authors.lastname: Nardone
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, 10032, USA.|Howard Hughes Medical Institute, Columbia University, New York, NY, 10032, USA.
authors.forename: Jeremie
authors.initials: J
authors.lastname: Vendome
authors.affiliation: School of Chemistry and Biomedical Sciences Research Centre, University of St. Andrews, North Haugh, St. Andrews, KY16 9ST, UK.
authors.forename: Nicholas James
authors.identifier: 0000-0003-0630-0138
authors.initials: NJ
authors.lastname: Westwood
authors.affiliation: Department of Microbial Natural Products, Helmholtz Institute for Pharmaceutical Research, Helmholtz Centre for Infection Research, Saarland University, 66123, Saarbrücken, Germany.
authors.forename: Rolf
authors.identifier: 0000-0002-1042-5665
authors.initials: R
authors.lastname: Müller
authors.affiliation: Workgroup Structural Biology of Biosynthetic Enzymes, Helmholtz Institute for Pharmaceutical Research, Helmholtz Centre for Infection Research, Saarland University, Universitätscampus E8 1, 66123, Saarbrücken, Germany.
authors.forename: Jesko
authors.identifier: 0000-0002-7153-1365
authors.initials: J
authors.lastname: Koehnke
mesh_terms: D018360:Crystallography, X-Ray; D007931:Leucyl Aminopeptidase / Q000378:metabolism*; D008958:Models, Molecular; D008968:Molecular Conformation; D010456:Peptides, Cyclic / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D010456:Peptides, Cyclic; C024712:bottromycin; D007931:Leucyl Aminopeptidase
keywords: BotP; RiPPs; biosynthesis; bottromycin; leucyl-aminopeptidases