caa a22 4500 475744144 CHVBK 20180406123508.0 cr unu---uuuuu 170329e20001001xx s 000 0 eng 10.1023/A:1007198202016 doi (NATIONALLICENCE)springer-10.1023/A:1007198202016 Characterization of Nicking of the Nontoxic-Nonhemagglutinin Components of Clostridium botulinum Types C and D Progenitor Toxin [Elektronische Daten] [Yoshimasa Sagane, Toshihiro Watanabe, Hirokazu Kouguchi, Hiroyuki Sunagawa, Kaoru Inoue, Yukako Fujinaga, Keiji Oguma, Tohru Ohyama] Clostridium botulinum C and D strains produce two types of progenitor toxins, M and L. Previously we reported that a 130-kDa nontoxic-nonhemagglutinin (NTNHA) component of the M toxin produced by type D strain CB16 was nicked at a unique site, leading to a 15-kDa N-terminal fragment and a 115-kDa C-terminal fragment. In this study, we identified the amino acid sequences around the nicking sites in the NTNHAs of the M toxins produced by C. botulinum type C and D strains by analysis of their C-terminal and N-terminal sequences and mass spectrometry. The C-terminus of the 15-kDa fragments was identified as Lys127 from these strains, indicating that a bacterial trypsin-like protease is responsible for the nicking. The 115-kDa fragment had mixtures of three different N-terminal amino acid sequences beginning with Leu135, Val139, and Ser141, indicating that 7-13 amino acid residues were deleted from the nicking site. The sequence beginning with Leu135 would also suggest cleavage by a trypsin-like protease, while the other two N-terminal amino acid sequences beginning with Val139 and Ser141 would imply proteolysis by an unknown protease. The nicked NTNHA forms a binary complex of two fragments that could not be separated without sodium dodecyl sulfate. Plenum Publishing Corporation, 2000 Clostridium botulinum nationallicence progenitor toxin nationallicence nontoxic-nonhemagglutinin nationallicence nicking nationallicence trypsin-like protease nationallicence Sagane Yoshimasa Department of Food Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, 099-2493, Abashiri, Japan aut Watanabe Toshihiro Department of Food Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, 099-2493, Abashiri, Japan aut Kouguchi Hirokazu Department of Food Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, 099-2493, Abashiri, Japan aut Sunagawa Hiroyuki Hokkaido Institute of Public Health, N19, W12, Kita-Ku, 060-0819, Sapporo, Japan aut Inoue Kaoru Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-Cho, 700-0914, Okayama, Japan aut Fujinaga Yukako Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-Cho, 700-0914, Okayama, Japan aut Oguma Keiji Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-Cho, 700-0914, Okayama, Japan aut Ohyama Tohru Department of Food Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, 099-2493, Abashiri, Japan aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/7(2000-10-01), 575-581 0277-8033 19:7<575 2000 19 10930 https://doi.org/10.1023/A:1007198202016 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1007198202016 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sagane Yoshimasa Department of Food Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, 099-2493, Abashiri, Japan aut NATIONALLICENCE 700 1- Watanabe Toshihiro Department of Food Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, 099-2493, Abashiri, Japan aut NATIONALLICENCE 700 1- Kouguchi Hirokazu Department of Food Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, 099-2493, Abashiri, Japan aut NATIONALLICENCE 700 1- Sunagawa Hiroyuki Hokkaido Institute of Public Health, N19, W12, Kita-Ku, 060-0819, Sapporo, Japan aut NATIONALLICENCE 700 1- Inoue Kaoru Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-Cho, 700-0914, Okayama, Japan aut NATIONALLICENCE 700 1- Fujinaga Yukako Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-Cho, 700-0914, Okayama, Japan aut NATIONALLICENCE 700 1- Oguma Keiji Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-Cho, 700-0914, Okayama, Japan aut NATIONALLICENCE 700 1- Ohyama Tohru Department of Food Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, 099-2493, Abashiri, Japan aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/7(2000-10-01), 575-581 0277-8033 19:7<575 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer