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Maturation and localization of the TolB protein required for colicin import.

By M Isnard, A Rigal, J C Lazzaroni, C Lazdunski and R Lloubes

Abstract

The tolB gene has been shown previously to encode two proteins of 47.5 kDa (TolB) and 43 kDa (TolB*). To explain the presence of these two forms, two hypotheses have been proposed: TolB might be posttranslationally processed to TolB*, or an internal in-frame translation initiation resulting in TolB* may occur (S. K. Levengood and R. E. Webster, J. Bacteriol. 171:6600-6609, 1989). To address this question, TolB was tagged by inserting in its C-terminal region an epitope recognized by monoclonal antibody 1C11 without altering the function of TolB. It was then demonstrated that the functional protein corresponded to TolB*, the mature periplasmic protein, and that TolB was its precursor form, which was observed only when the protein was overexpressed. These two forms were purified by immunoprecipitation, and their N-terminal sequences were determined. An antibody directed against TolB was raised, which confirmed the results obtained with the tagged TolB

Topics: Research Article
Year: 1994
DOI identifier: 10.1128/jb.176.20.6392-6396.1994
OAI identifier: oai:pubmedcentral.nih.gov:196981
Provided by: PubMed Central
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