Vibrio spp. secrete proaerolysin as a folded dimer without the need for disulphide bond formation

Molecular Microbiology
K R HardieJ T Buckley

Abstract

Proaerolysin is an extracellular dimeric protein that is secreted across the inner and outer membranes of Aeromonas spp. in separate steps. To investigate the role of protein folding in the second step, one or more cysteine residues were introduced and the mutant proaerolysins were expressed in Aeromonas hydrophila and Aeromonas salmonicida, as well as Vibrio cholerae. Replacing Met-41 with Cys resulted in expression of a protein that could form a dimer in which the monomers were linked together by a disulphide bridge. A double mutant was also made, in which Gly-202 and Ile-445 were replaced with cysteine in order to allow the formation of an intrachain disulphide bridge when the molecule was correctly folded. The M41C covalent dimer and G202C/I445C proaerolysin with the new intrachain bridge were both easily detected inside the bacteria, and they later appeared in the culture supernatants. Small amounts of incorrectly folded proaerolysin were also observed in the cells, but they were not secreted. We observed in the cells, but they were not secreted. We conclude that proaerolysin folds and dimerizes before being released from the cell, and that correct folding is a requirement for secretion to occur. The proton ionophore CCCP ...Continue Reading

Citations

Oct 12, 2001·Toxicon : Official Journal of the International Society on Toxinology·M FivazF G van der Goot
Apr 3, 2012·Nature Reviews. Microbiology·Konstantin V KorotkovWim G J Hol
May 1, 1997·Molecular Biology of the Cell·S A Clark, S M Theg
Jun 1, 1997·The EMBO Journal·J RossjohnM W Parker
Sep 5, 2001·The EMBO Journal·L Debarbieux, C Wandersman
Mar 14, 2012·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Badreddine DouziRomé Voulhoux
Jan 3, 2001·Journal of Bacteriology·A UrbanK E Jaeger
Sep 29, 2011·Canadian Journal of Microbiology·Fang BaiMingqiang Qiao
Aug 8, 2014·Molecular Microbiology·Camille PineauVladimir E Shevchik
Nov 18, 2004·Biochimica Et Biophysica Acta·Alain Filloux
Dec 14, 1999·The Journal of Biological Chemistry·C LesieurF G van der Goot
Dec 23, 1999·The Journal of Biological Chemistry·M FivazF G van der Goot
Nov 26, 1998·The Journal of Biological Chemistry·L AbramiF G van der Goot
May 15, 2001·The Journal of Biological Chemistry·P BraunJ Tommassen
Apr 20, 2001·Molecular Microbiology·M Sandkvist
Aug 14, 2003·FEBS Letters·Silvia E T BittencourtMarcelo V Sousa
May 10, 2017·Molecular Microbiology·Jenny-Lee ThomassinOlivera Francetic
Jan 7, 2000·Microbial Pathogenesis·T NomuraK Okamoto
Apr 1, 1996·Journal of Bacteriology·M J Brumlik, J T Buckley
Feb 16, 2019·EcoSal Plus·Konstantin V Korotkov, Maria Sandkvist

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