Hydrophobic surface patches on LolA of Pseudomonas aeruginosa are essential for lipoprotein binding.

TitleHydrophobic surface patches on LolA of Pseudomonas aeruginosa are essential for lipoprotein binding.
Publication TypeJournal Article
Year of Publication2010
AuthorsRemans, K., K. Pauwels, P. van Ulsen, L. Buts, P. Cornelis, J. Tommassen, S. N. Savvides, K. Decanniere, and P. Van Gelder
JournalJ Mol Biol
Volume401
Issue5
Pagination921-30
Date Published2010 Sep 3
ISSN1089-8638
KeywordsBacterial Proteins, Base Sequence, Circular Dichroism, Cloning, Molecular, Crystallization, Crystallography, X-Ray, DNA Primers, Electrophoresis, Polyacrylamide Gel, Lipoproteins, Models, Molecular, Protein Binding, Protein Conformation, Pseudomonas aeruginosa, Surface Properties
Abstract

Many lipoproteins reside in the outer membrane (OM) of Gram-negative bacteria, and their biogenesis is dependent on the Lol (localization of lipoproteins) system. The periplasmic chaperone LolA accepts OM-destined lipoproteins that are released from the inner membrane by the LolCDE complex and transfers them to the OM receptor LolB. The exact nature of the LolA-lipoprotein complex is still unknown. The crystal structure of Escherichia coli LolA features an open beta-barrel covered by alpha helices that together constitute a hydrophobic cavity, which would allow the binding of one acyl chain. However, OM lipoproteins contain three acyl chains, and the stoichiometry of the LolA-lipoprotein complex is 1:1. Here we present the crystal structure of Pseudomonas aeruginosa LolA that projects clear hydrophobic surface patches. Since these patches are large enough to accommodate acyl chains, their role in lipoprotein binding was investigated. Several LolA mutant proteins were created, and their functionality was assessed by studying their capacity to release lipoproteins produced in sphaeroplasts. Interruption of the largest hydrophobic patch completely destroyed the lipoprotein-releasing capacity of LolA, while interruption of smaller patches apparently reduced efficiency. Thus, the results show a new lipoprotein transport model that places (some of) the acyl chains on the hydrophobic surface patches.

DOI10.1016/j.jmb.2010.06.067
Alternate JournalJ. Mol. Biol.
PubMed ID20620146