Structural and functional significance of the FGL sequence of the periplasmic chaperone Caf1M of Yersinia pestis.

Abstract

The periplasmic molecular chaperone Caf1M of Yersinia pestis is a typical representative of a subfamily of specific chaperones involved in assembly of surface adhesins with a very simple structure. One characteristic feature of this Caf1M-like subfamily is possession of an extended, variable sequence (termed FGL) between the F1 and subunit binding G1 beta-strands. In contrast, FGS subfamily members, characterized by PapD, have a short F1-G1 loop and are involved in assembly of complex pili. To elucidate the structural and functional significance of the FGL sequence, a mutant Caf1M molecule (dCaf1M), in which the 27 amino acid residues between the F1 and G1 beta-strands had been deleted, was constructed. Expression of the mutated caf1M in Escherichia coli resulted in accumulation of high levels of dCaf1M. The far-UV circular dichroism spectra of the mutant and wild-type proteins were indistinguishable and exhibited practically the same temperature and pH dependencies. Thus, the FGL sequence of Caf1M clearly does not contribute significantly to the stability of the protein conformation. Preferential cleavage of Caf1M by trypsin at Lys-119 confirmed surface exposure of this part of the FGL sequence in the isolated chaperone and periplasmic chaperone-subunit complex. There was no evidence of surface-localized Caf1 subunit in the presence of the Caf1A outer membrane protein and dCaf1M. In contrast to Caf1M, dCaf1M was not able to form a stable complex with Caf1 nor could it protect the subunit from proteolytic degradation in vivo. This demonstration that the FGL sequence is required for stable chaperone-subunit interaction, but not for folding of a stable chaperone, provides a sound basis for future detailed molecular analyses of the FGL subfamily of chaperones.

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Cite this paper

@article{Chapman1999StructuralAF, title={Structural and functional significance of the FGL sequence of the periplasmic chaperone Caf1M of Yersinia pestis.}, author={David Alan Chapman and Anton V. Zavialov and T. V. Chernovskaya and Andrey V Karlyshev and Galina A Zav'yalova and Anatoly M Vasiliev and I. V. Dudich and Vyacheslav M. Abramov and Vladimir P. Zav'yalov and Stuart Macintyre}, journal={Journal of bacteriology}, year={1999}, volume={181 8}, pages={2422-9} }