title: Identification and Characterization of a New Enterotoxin Produced by Clostridium perfringens Isolated from Food Poisoning Outbreaks.
abstract: There is a strain of Clostridium perfringens, W5052, which does not produce a known enterotoxin. We herein report that the strain W5052 expressed a homologue of the iota-like toxin components sa and sb of C. spiroforme, named Clostridium perfringens iota-like enterotoxin, CPILE-a and CPILE-b, respectively, based on the results of a genome sequencing analysis and a systematic protein screening. In the nicotinamide glyco-hydrolase (NADase) assay the hydrolysis activity was dose-dependently increased by the concentration of rCPILE-a, as judged by the mass spectrometry analysis. In addition, the actin monomer of the lysates of Vero and L929 cells were radiolabeled in the presence of [32P]NAD and rCPILE-a. These findings indicated that CPILE-a possesses ADP-ribosylation activity. The culture supernatant of W5052 facilitated the rounding and killing of Vero and L929 cells, but the rCPILE-a or a non-proteolyzed rCPILE-b did not. However, a trypsin-treated rCPILE-b did. Moreover, a mixture of rCPILE-a and the trypsin-treated rCPILE-b enhanced the cell rounding and killing activities, compared with that induced by the trypsin-treated rCPILE-b alone. The injection of the mixture of rCPILE-a and the trypsin-treated rCPILE-b into an ileum loop of rabbits evoked the swelling of the loop and accumulation of the fluid dose-dependently, suggesting that CPILE possesses enterotoxic activity. The evidence presented in this communication will facilitate the epidemiological, etiological, and toxicological studies of C. perfringens food poisoning, and also stimulate studies on the transfer of the toxins' gene(s) among the Genus Clostridium.
authors.affiliation: Division of Microbiology, National Institute of Health Sciences, Tokyo, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Irikura
authors.affiliation: Department of Microbiology, Tokyo Metropolitan Institute of Public Health, Tokyo, Japan.
authors.forename: Chie
authors.initials: C
authors.lastname: Monma
authors.affiliation: Department of Microbiology, Tokyo Metropolitan Institute of Public Health, Tokyo, Japan.
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Suzuki
authors.affiliation: Department of Microbiology, Tokyo Metropolitan Institute of Public Health, Tokyo, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Nakama
authors.affiliation: Department of Microbiology, Tokyo Metropolitan Institute of Public Health, Tokyo, Japan.
authors.forename: Akemi
authors.initials: A
authors.lastname: Kai
authors.affiliation: Department of Molecular Biology, Research Institute of Microbial Diseases, Osaka University, Osaka, Japan.
authors.forename: Aya
authors.initials: A
authors.lastname: Fukui-Miyazaki
authors.affiliation: Department of Molecular Biology, Research Institute of Microbial Diseases, Osaka University, Osaka, Japan.
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Horiguchi
authors.affiliation: Division of Microbiology, National Institute of Health Sciences, Tokyo, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Yoshinari
authors.affiliation: Division of Microbiology, National Institute of Health Sciences, Tokyo, Japan.|Department of Food and Life Sciences, The Graduate School of Life and Environmental Sciences, Azabu University, Kanagawa, Japan.
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Sugita-Konishi
authors.affiliation: Division of Microbiology, National Institute of Health Sciences, Tokyo, Japan.|Department of Veterinary Medicine, Iwate University, Morioka, Japan.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Kamata
mesh_terms: D020727:ADP-Ribosylation Factors / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D003015:Clostridium Infections / Q000453:epidemiology / Q000382:microbiology*; D003016:Clostridium perfringens / Q000235:genetics* / Q000302:isolation & purification; D017124:Conserved Sequence; D004196:Disease Outbreaks; D004768:Enterotoxins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D005517:Foodborne Diseases / Q000453:epidemiology / Q000382:microbiology*; D015870:Gene Expression; D006801:Humans; D007082:Ileum / Q000382:microbiology; D008297:Male; D008969:Molecular Sequence Data; D009244:NAD+ Nucleosidase / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D011817:Rabbits; D017422:Sequence Analysis, DNA; D014041:Tokyo; D014709:Vero Cells
chemical_list: D004768:Enterotoxins; D009244:NAD+ Nucleosidase; D020727:ADP-Ribosylation Factors