title: Structural analysis of leader peptide binding enables leader-free cyanobactin processing.
abstract: Regioselective modification of amino acids within the context of a peptide is common to a number of biosynthetic pathways, and many of the resulting products have potential as therapeutics. The ATP-dependent enzyme LynD heterocyclizes multiple cysteine residues to thiazolines within a peptide substrate. The enzyme requires the substrate to have a conserved N-terminal leader for full activity. Catalysis is almost insensitive to immediately flanking residues in the substrate, suggesting that recognition occurs distant from the active site. Nucleotide and peptide substrate co-complex structures of LynD reveal that the substrate leader peptide binds to and extends the β-sheet of a conserved domain of LynD, whereas catalysis is accomplished in another conserved domain. The spatial segregation of catalysis from recognition combines seemingly contradictory properties of regioselectivity and promiscuity, and it appears to be a conserved strategy in other peptide-modifying enzymes. A variant of LynD that efficiently processes substrates without a leader peptide has been engineered.
authors.affiliation: BSRC, University of St Andrews, St Andrews, KY16 9RH.
authors.forename: Jesko
authors.initials: J
authors.lastname: Koehnke
authors.affiliation: BSRC, University of St Andrews, St Andrews, KY16 9RH.
authors.forename: Greg
authors.initials: G
authors.lastname: Mann
authors.affiliation: BSRC, University of St Andrews, St Andrews, KY16 9RH.
authors.forename: Andrew F
authors.initials: AF
authors.lastname: Bent
authors.affiliation: BSRC, University of St Andrews, St Andrews, KY16 9RH.
authors.forename: Hannes
authors.initials: H
authors.lastname: Ludewig
authors.affiliation: BSRC, University of St Andrews, St Andrews, KY16 9RH.
authors.forename: Sally
authors.initials: S
authors.lastname: Shirran
authors.affiliation: BSRC, University of St Andrews, St Andrews, KY16 9RH.
authors.forename: Catherine
authors.initials: C
authors.lastname: Botting
authors.affiliation: BSRC, University of St Andrews, St Andrews, KY16 9RH.
authors.forename: Tomas
authors.initials: T
authors.lastname: Lebl
authors.affiliation: Marine Biodiscovery Centre, Department of Chemistry, University of Aberdeen, Meston Walk, Aberdeen, AB24 3UE.|Institute of Medical Sciences, University of Aberdeen, Aberdeen AB25 2ZD, Scotland, UK.|Pharmacognosy Department, Faculty of Pharmacy, Mansoura University, Mansoura 35116, Egypt.
authors.forename: Wael
authors.initials: W
authors.lastname: Houssen
authors.affiliation: Marine Biodiscovery Centre, Department of Chemistry, University of Aberdeen, Meston Walk, Aberdeen, AB24 3UE.
authors.forename: Marcel
authors.initials: M
authors.lastname: Jaspars
authors.affiliation: BSRC, University of St Andrews, St Andrews, KY16 9RH.|State Key Laboratory of Biotherapy and Collaborative Innovation Center for Biotherapy, West China Hospital, Sichuan University, China.
authors.forename: James H
authors.initials: JH
authors.lastname: Naismith
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D055162:Biocatalysis; D000458:Cyanobacteria / Q000737:chemistry / Q000378:metabolism; D003500:Cyclization; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D015870:Gene Expression; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity; D013844:Thiazoles / Q000737:chemistry / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D010456:Peptides, Cyclic; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D013844:Thiazoles; D000255:Adenosine Triphosphate; D003545:Cysteine