PMID:7929011
| Citation |
Isnard, M, Rigal, A, Lazzaroni, JC, Lazdunski, C and Lloubes, R (1994) Maturation and localization of the TolB protein required for colicin import. J. Bacteriol. 176:6392-6 |
|---|---|
| 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. |
| Links | |
| Keywords |
Amino Acid Sequence; Bacterial Proteins/biosynthesis; Bacterial Proteins/chemistry; Bacterial Proteins/genetics; Base Sequence; Biological Transport; Cell Compartmentation; Colicins/metabolism; Escherichia coli Proteins; Gene Expression Regulation, Bacterial; Membrane Proteins/biosynthesis; Membrane Proteins/chemistry; Membrane Proteins/genetics; Molecular Sequence Data; Periplasmic Proteins; Protein Precursors/chemistry; Protein Precursors/genetics; Protein Precursors/metabolism; Protein Processing, Post-Translational; Sequence Analysis |
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