Comparison of type 5d autotransporter phospholipases demonstrates a correlation between high activity and intracellular pathogenic lifestyle

Trunk, T., Casasanta, M.A., Yoo, C.C., Slade, D.J. and Leo, J.C. ORCID: 0000-0002-7066-7527, 2019. Comparison of type 5d autotransporter phospholipases demonstrates a correlation between high activity and intracellular pathogenic lifestyle. Biochemical Journal. ISSN 0264-6021

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Abstract

Autotransporters, or type 5 secretion systems, are widespread surface proteins of Gram-negative bacteria often associated with virulence functions. Autotransporters consist of an outer membrane β-barrel domain and an exported passenger. In the poorly studied type 5d subclass, the passenger is a patatin-like lipase. The prototype of this secretion pathway is PlpD of Pseudomonas aeruginosa , an opportunistic human pathogen. The PlpD passenger is a homodimer with phospholipase A1 (PLA1) activity. Based on sequencing data, PlpD-like proteins are present in many bacterial species. We characterized the enzymatic activity, specific lipid binding and oligomeric status of PlpD homologs from Aeromonas hydrophila (a fish pathogen), Burkholderia pseudomallei (a human pathogen) and Ralstonia solanacearum (a plant pathogen) and compared these with PlpD. We demonstrate that recombinant type 5d-secreted patatin domains have lipase activity and form dimers or higher-order oligomers. However, dimerization is not necessary for lipase activity; in fact, by making monomeric variants of PlpD, we show that enzymatic activity slightly increases while protein stability decreases. The lipases from the intracellular pathogens A. hydrophila and B. pseudomallei display PLA2 activity in addition to PLA1 activity. Although the type 5d-secreted lipases from the animal pathogens bound to intracellular lipid targets, phosphatidylserine and phosphatidylinositol phosphates, hydrolysis of these lipids could only be observed for FplA of Fusobacterium nucleatum . Yet, we noted a correlation between high lipase activity in type 5d autotransporters and intracellular lifestyle. We hypothesize that type 5d phospholipases are intracellularly active and function in modulation of host cell signaling events.

Item Type: Journal article
Publication Title: Biochemical Journal
Creators: Trunk, T., Casasanta, M.A., Yoo, C.C., Slade, D.J. and Leo, J.C.
Publisher: Portland Press
Date: 6 September 2019
ISSN: 0264-6021
Identifiers:
NumberType
10.1042/bcj20190136DOI
Divisions: Schools > School of Science and Technology
Record created by: Jonathan Gallacher
Date Added: 16 Sep 2019 10:45
Last Modified: 06 Sep 2020 03:00
URI: http://irep.ntu.ac.uk/id/eprint/37668

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