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Structure. 2015 Apr 7;23(4):762-73. doi: 10.1016/j.str.2015.01.022. Epub 2015 Mar 19.

Probing the protein interaction network of Pseudomonas aeruginosa cells by chemical cross-linking mass spectrometry.

Author information

1
Department of Genome Sciences, University of Washington, 850 Republican Street, Seattle, WA 98109, USA.
2
Department of Microbiology, University of Washington Medical Center, 1959 N.E. Pacific Street, Seattle, WA 98195, USA.
3
Department of Genome Sciences, University of Washington, 850 Republican Street, Seattle, WA 98109, USA. Electronic address: jimbruce@uw.edu.

Abstract

In pathogenic Gram-negative bacteria, interactions among membrane proteins are key mediators of host cell attachment, invasion, pathogenesis, and antibiotic resistance. Membrane protein interactions are highly dependent upon local properties and environment, warranting direct measurements on native protein complex structures as they exist in cells. Here we apply in vivo chemical cross-linking mass spectrometry, to reveal the first large-scale protein interaction network in Pseudomonas aeruginosa, an opportunistic human pathogen, by covalently linking interacting protein partners, thereby fixing protein complexes in vivo. A total of 626 cross-linked peptide pairs, including previously unknown interactions of many membrane proteins, are reported. These pairs not only define the existence of these interactions in cells but also provide linkage constraints for complex structure predictions. Structures of three membrane proteins, namely, SecD-SecF, OprF, and OprI are predicted using in vivo cross-linked sites. These findings improve understanding of membrane protein interactions and structures in cells.

PMID:
25800553
PMCID:
PMC4756656
DOI:
10.1016/j.str.2015.01.022
[Indexed for MEDLINE]
Free PMC Article

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