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Researchers narrow search for structure of cholera toxins extracellular transport signal

BUFFALO, N.Y. -- Working with Vibrio cholerae, the bacterium that causes the severe diarrheal disease of cholera, microbiologists at the University at Buffalo have revealed new information on a cellular signaling system that ultimately will help scientists understand how cholera toxin and virulent proteins of other pathogenic bacteria migrate through their cellular membranes to cause disease.

Using chitinase, a protein known to be secreted by V. cholerae by the same mechanism as cholera toxin, the researchers engineered a series of insertions, deletions and mutations in its amino-acid chain. Using this mutagenic approach, they determined that the extracellular transport signal of chitinase was encoded by amino acids located between positions 75 and 555 on the chain. Further experiments demonstrated that only a portion of this 480-amino-acid-region was essential for secretion of chitinase.

"In addition to providing new information about the transport of chitinase and cholera toxin, these findings increase our basic understanding of the methods by which proteins in general are transported across membranes, an essential activity for any living cell," said Terry D. Connell, Ph.D., associate professor of microbiology in the UB School of Medicine and Biomedical Sciences and senior author on the research.

Results of the research appear in the April issue (Vol. 184, No. 8) of the Journal of Bacteriology.

Cholera toxin is known to be secreted from the bacterial cell by a complex secretory machinery. However, rather than concentrating on the secretory mechanism, a focus of several laboratories at other institutions, Connell and colleagues Jason Folster, a doctoral student, and Daniel Metzger, UB research associate, working in the university's Witebsky Center for Microbial Pathogenesis and Immunology, set out to investigate the structural signals on cholera toxin that initiate its translocation.

The UB scientists are the
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Contact: Lois Baker
ljbaker@buffalo.edu
716-645-5000 x1417
University at Buffalo
27-Mar-2002


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