title: Interaction of FkpA, a peptidyl-prolyl cis/trans isomerase with EspP autotransporter protein.
abstract: The serine protease autotransporters of Enterobacteriaceae (SPATEs) represent a large class of proteases with contributions to virulence. They are synthesized with a C-terminal domain that forms a β-barrel pore in the outer membrane implicated in translocation of the N-terminal 'passenger' domain across the outer membrane. The most recent model for autotransporter secretion comprises entry to the periplasm via the Sec apparatus, followed by the insertion of the C-terminus into the outer membrane as a β-barrel protein and accompanied by translocation of the passenger domain to the bacterial cell surface, all of this with the assistance of the Bam complex insertase/foldase and periplasmic chaperone proteins. We have recently observed direct involvement of periplasmic chaperones in the biogenesis of EspP, a prototypical autotransporter protein produced by Escherichia coli O157:H7. Using molecular and biophysical approaches we demonstrated for the first time, direct protein-protein interactions between the periplasmic SurA and DegP chaperones and either the EspP-β or EspP passenger domains. Such chaperone interactions took place on conserved aromatic residues on the SPATE family. In this report, we now demonstrate direct binding of the periplasmic chaperone FkpA to the EspP passanger domain in Surface Plasmon Resonance experiments with relatively high affinity. We also provide evidence of interaction between the SurA and Skp chaperones with the Bam. These findings in conjunction with newly published data support the role of chaperones in preventing misfolding of AT passenger domains before translocation throughout the Bam complex.
authors.affiliation: Center for Vaccine Development; University of Maryland School of Medicine; Baltimore, MD USA.
authors.forename: Fernando
authors.initials: F
authors.lastname: Ruiz-Perez
authors.forename: Ian R
authors.initials: IR
authors.lastname: Henderson
authors.forename: James P
authors.initials: JP
authors.lastname: Nataro