Incorporation of a polypeptide segment into the beta-domain pore during the assembly of a bacterial autotransporter.

Abstract:

:Bacterial autotransporters consist of an N-terminal 'passenger domain' that is transported into the extracellular space by an unknown mechanism and a C-terminal 'beta-domain' that forms a beta-barrel in the outer membrane. Recent studies have revealed that fully assembled autotransporters have an unusual architecture in which a small passenger domain segment traverses the pore formed by the beta-domain. It is unclear, however, whether this configuration forms prior to passenger domain translocation or results from the translocation of the passenger domain through the beta-domain pore. By examining the accessibility of tobacco etch virus protease sites and single-cysteine residues in the passenger domain of the Escherichia coli O157:H7 autotransporter EspP at different stages of protein biogenesis, we identified a novel pre-translocation intermediate whose topology resembles that of the fully assembled protein. This intermediate was isolated in the periplasm in cell fractionation experiments. The data strongly suggest that the EspP beta-domain and an embedded polypeptide segment are integrated into the outer membrane as a single pre-formed unit. The data also provide indirect evidence that at least some outer membrane proteins acquire considerable tertiary structure prior to their membrane integration.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Ieva R,Skillman KM,Bernstein HD

doi

10.1111/j.1365-2958.2007.06048.x

subject

Has Abstract

pub_date

2008-01-01 00:00:00

pages

188-201

issue

1

eissn

0950-382X

issn

1365-2958

pii

MMI6048

journal_volume

67

pub_type

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