title: Chlorinated Glycopeptide Antibiotic Peptide Precursors Improve Cytochrome P450-Catalyzed Cyclization Cascade Efficiency.
abstract: The activity of glycopeptide antibiotics (GPAs) depends upon important structural modifications to their precursor heptapeptide backbone: specifically, the cytochrome P450-catalyzed oxidative cross-linking of aromatic side chains as well as the halogenation of specific residues within the peptide. The timing of halogenation and its effect on the cyclization of the peptide are currently unclear. Our results show that chlorination of peptide precursors improves their processing by P450 enzymes in vitro, which provides support for GPA halogenation occurring prior to peptide cyclization during nonribosomal peptide synthesis. We could also determine that the activity of the second enzyme in the oxidative cyclization cascade, OxyA, remains higher for chlorinated peptide substrates even when the biosynthetic GPA product possesses an altered chlorination pattern, which supports the role of the chlorine atoms in orienting the peptide substrate in the active site of these enzymes.
authors.affiliation: Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research , Jahnstrasse 29, 69120 Heidelberg, Germany.
authors.forename: Madeleine
authors.initials: M
authors.lastname: Peschke
authors.affiliation: Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research , Jahnstrasse 29, 69120 Heidelberg, Germany.
authors.forename: Clara
authors.initials: C
authors.lastname: Brieke
authors.affiliation: Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University , Clayton, Victoria 3800, Australia.|Monash Biomedical Proteomics Facility, Monash University , Clayton, Victoria 3800, Australia.
authors.forename: Rob J A
authors.initials: RJ
authors.lastname: Goode
authors.affiliation: Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University , Clayton, Victoria 3800, Australia.|Monash Biomedical Proteomics Facility, Monash University , Clayton, Victoria 3800, Australia.
authors.forename: Ralf B
authors.initials: RB
authors.lastname: Schittenhelm
authors.affiliation: Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research , Jahnstrasse 29, 69120 Heidelberg, Germany.|Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University , Clayton, Victoria 3800, Australia.|ARC Centre of Excellence in Advanced Molecular Imaging, Monash University , Clayton, Victoria 3800, Australia.|EMBL Australia, Monash University , Clayton, Victoria 3800, Australia.
authors.forename: Max J
authors.identifier: 0000-0002-9739-6157
authors.initials: MJ
authors.lastname: Cryle
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000494:pharmacology*; D055162:Biocatalysis / Q000187:drug effects*; D020134:Catalytic Domain; D003500:Cyclization / Q000187:drug effects; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000378:metabolism*; D006020:Glycopeptides / Q000737:chemistry* / Q000494:pharmacology*; D054879:Halogenation; D010084:Oxidation-Reduction
chemical_list: D000900:Anti-Bacterial Agents; D006020:Glycopeptides; D003577:Cytochrome P-450 Enzyme System