Structural basis for alginate secretion across the bacterial outer membrane

John C Whitney, Iain Hay, Canhui Danny Li, Paul D W Eckford, Howard Robinson, Maria F Amaya, Lynn F Wood, Dennis E Ohman, Christine E Bear, Bernd H A Rehm, P Lynne Howell

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54 Citations (Scopus)

Abstract

Pseudomonas aeruginosa is the predominant pathogen associated with chronic lung infection among cystic fibrosis patients. During colonization of the lung, P. aeruginosa converts to a mucoid phenotype characterized by the overproduction of the exopolysaccharide alginate. Secretion of newly synthesized alginate across the outer membrane is believed to occur through the outer membrane protein AlgE. Here we report the 2.3 ? crystal structure of AlgE, which reveals a monomeric 18-stranded ?-barrel characterized by a highly electropositive pore constriction formed by an argininerich conduit that likely acts as a selectivity filter for the negatively charged alginate polymer. Interestingly, the pore constriction is occluded on either side by extracellular loop L2 and an unusually long periplasmic loop, T8. In halide efflux assays, deletion of loop T8 (?T8-AlgE) resulted in a threefold increase in anion flux compared to the wild-type or ?L2-AlgE supporting the idea that AlgE forms a transport pathway through the membrane and suggesting that transport is regulated by T8. This model is further supported by in vivo experiments showing that complementation of an algE deletion mutant with ?T8-AlgE impairs alginate production. Taken together, these studies support a mechanism for exopolysaccharide export across the outer membrane that is distinct from the Wza-mediated translocation observed in canonical capsular polysaccharide export systems.
Original languageEnglish
Pages (from-to)13083 - 13088
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume108
Issue number32
DOIs
Publication statusPublished - 2011
Externally publishedYes

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