Isolation and characterization of a cytotoxin produced by Plesiomonas shigelloides P-1 strain

Yoshio Okawa, Yuko Ohtomo, Hitoshi Tsugawa, Yuko Matsuda, Hidemitsu Kobayashi, Teizo Tsukamoto

研究成果: Article査読

26 被引用数 (Scopus)

抄録

In order to clarify the enteropathogenicity of Plesiomonas shigelloides, we investigated a cytotoxin produced by the P-1 strain isolated from patients suffering from diarrhea. The cytotoxicity of the culture filtrate of the strain reached a maximum in culture at 37°C after 12 h shaken in BHI medium. The cytotoxin in the cultures was purified by (NH 4) 2SO 4 precipitation, and Sephacryl S-100, Mono Q HR, and Superdex 200 HR column chromatographies. An approximate 340-fold purification was achieved, with a recovery of about 1.4%, from the culture supernatant. The cytotoxin is heat-stable, and is a complex of three major proteins (LPS-binding proteins with molecular weights of 32, 40, and 48 kDa), with lipopolysaccharide (LPS) giving a total a molecular weight of more than 600 kDa. The ratio of protein to LPS in the cytotoxin was 6-5. The cytotoxic activity was reduced by about 80% by proteinase K treatment or when incubated with anti-cholera toxin antibody (Anti-CT). Western blotting of the cytotoxin with Anti-CT demonstrated the presence of two anti-cholera toxin-reactive protein (ACRP) bands with molecular weights of 40 kDa (a major single protein band) and 48 kDa. The N-terminal amino acid sequence (20 residues) of the 40 kDa protein was 75% identical to Pasteurella multocida cell membrane proteins. The cytotoxin gave a positive reaction in the suckling mouse assay whereas LPS alone hardly exhibited any cytotoxic or enterotoxigenic activity. In conclusion, P. shigelloides produces a cytotoxin that consists of a complex of protein and LPS with the former component exhibiting both cytotoxicity and enteropathogenicity. This cytotoxin has the potential to have an important role in the enteropathogenicity of P. shigelloides.

本文言語English
ページ(範囲)125-130
ページ数6
ジャーナルFEMS microbiology letters
239
1
DOI
出版ステータスPublished - 2004 10 1
外部発表はい

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Genetics

フィンガープリント 「Isolation and characterization of a cytotoxin produced by Plesiomonas shigelloides P-1 strain」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル