Document Detail

Preliminary evidence that Clostridium perfringens type A enterotoxin is present in a 160,000-Mr complex in mammalian membranes.
MedLine Citation:
PMID:  2536357     Owner:  NLM     Status:  MEDLINE    
Clostridium perfringens type A 125I-enterotoxin (125I-CPE) was bound to rabbit intestinal brush border membranes (BBMs) or Vero cells and then solubilized with 3-[(3-cholamidopropyl)dimethyl-ammonio]-1-propanesulfonate (CHAPS). Solubilized radioactivity was analyzed by gel filtration chromatography on a Sepharose 4B column or by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) without sample boiling and autoradiography. Specifically bound 125I-CPE extracted from either BBMs or Vero cells was primarily associated with a complex of approximately 160,000 Mr. The CPE complex was partially purified by gel filtration or SDS-PAGE without sample boiling. SDS-PAGE analysis with sample boiling of the partially purified 125I-CPE complex from Vero cells or BBMs suggested that CPE complex contains both a 50,000-Mr protein and a 70,000-Mr protein in approximately equimolar amounts. This result is supported by affinity chromatography with CPE immobilized on Sepharose 4B, which showed the specific interaction of similar size proteins with CPE. The simplest explanation for these results is that CPE (Mr 35,000) interacts with 50,000-Mr and 70,000-Mr eucaryotic proteins to form a membrane-dependent complex of approximately 160,000 Mr. These results suggest that the receptor or target site(s) or both for CPE are similar in both BBMs and Vero cells. The significance of these findings in terms of CPE binding, insertion, and biologic action is discussed.
A P Wnek; B A McClane
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Infection and immunity     Volume:  57     ISSN:  0019-9567     ISO Abbreviation:  Infect. Immun.     Publication Date:  1989 Feb 
Date Detail:
Created Date:  1989-03-01     Completed Date:  1989-03-01     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0246127     Medline TA:  Infect Immun     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  574-81     Citation Subset:  IM    
Department of Microbiology, Biochemistry and Molecular Biology, University of Pittsburgh School of Medicine, Pennsylvania 15261.
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MeSH Terms
Bacterial Toxins / biosynthesis,  isolation & purification*
Calcium-Binding Proteins*
Cations, Divalent
Cell Membrane / analysis*,  metabolism,  microbiology
Cercopithecus aethiops
Chromatography, Affinity
Clostridium perfringens / analysis*,  metabolism
Electrophoresis, Polyacrylamide Gel
Macromolecular Substances
Microvilli / analysis,  microbiology
Molecular Weight
Type C Phospholipases*
Vero Cells
Grant Support
Reg. No./Substance:
0/Bacterial Toxins; 0/Calcium-Binding Proteins; 0/Cations, Divalent; 0/Macromolecular Substances; EC 3.1.4.-/Type C Phospholipases; EC toxin, Clostridium perfringens

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