title: Molecular characterization of the protease from Clostridium botulinum serotype C responsible for nicking in botulinum neurotoxin complex.
abstract: A protease was purified from the culture medium of Clostridium botulinum serotype C strain Stockholm (C-St). The purified protease belonged to the cysteine protease family based on assays for enzyme inhibitors, activators and kinetic parameters. The protease formed a binary complex consisting of 41- and 17-kDa proteins held together non-covalently. The DNA sequence encoding the protease gene was shown to be a single open reading frame of 1593 nucleotides, predicting 530 amino acid residues including a signal peptide. The N-terminal region of the native enzyme underwent further proteolytic modification after processing by a signal peptidase. The protease introduced intermolecular cleavage into an intact single chain botulinum neurotoxin (BoNT) at a specific site. Homology modeling and docking simulation of C-St BoNT and C-St protease demonstrated that the specific nicking-site of the BoNT appears to fit into the deep pocket in the active site of the protease.
authors.affiliation: Department of Medicinal and Life Science, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda 278-8510, Japan.
authors.forename: Tomonori
authors.initials: T
authors.lastname: Suzuki
authors.forename: Tohru
authors.initials: T
authors.lastname: Yoneyama
authors.forename: Keita
authors.initials: K
authors.lastname: Miyata
authors.forename: Akifumi
authors.initials: A
authors.lastname: Mikami
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Chikai
authors.forename: Ken
authors.initials: K
authors.lastname: Inui
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kouguchi
authors.forename: Koichi
authors.initials: K
authors.lastname: Niwa
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Watanabe
authors.forename: Satoru
authors.initials: S
authors.lastname: Miyazaki
authors.forename: Tohru
authors.initials: T
authors.lastname: Ohyama
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001905:Botulinum Toxins / Q000737:chemistry / Q000378:metabolism*; D002413:Cations, Divalent / Q000737:chemistry / Q000378:metabolism; D003014:Clostridium botulinum / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D003198:Computer Simulation; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D008670:Metals / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012703:Serotyping; D013379:Substrate Specificity
chemical_list: D002413:Cations, Divalent; D015853:Cysteine Proteinase Inhibitors; D008670:Metals; D003546:Cysteine Endopeptidases; D001905:Botulinum Toxins