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Biochem J. 2018 Jun 6;475(11):1885-1907. doi: 10.1042/BCJ20180257.

A phosphatidic acid-binding protein is important for lipid homeostasis and adaptation to anaerobic biofilm conditions in Pseudomonas aeruginosa.

Author information

1
Structure and Function of Proteins, Helmholtz Centre for Infection Research, Inhoffenstraße 7, D-38124 Braunschweig, Germany.
2
Institute of Microbiology, Technische Universität Braunschweig, Spielmannstraße 7, D-38106 Braunschweig, Germany.
3
Department of Microbial Biology, Ruhr University Bochum, D-44801 Bochum, Germany.
4
Cellular Proteome Research, Helmholtz Centre for Infection Research, Inhoffenstraße 7, D-38124 Braunschweig, Germany.
5
Institute of Microbiology, Technische Universität Braunschweig, Spielmannstraße 7, D-38106 Braunschweig, Germany j.moser@tu-bs.de.

Abstract

A quantitative Pseudomonas aeruginosa proteomics approach revealed increased abundance of the so-far uncharacterized protein PA3911 in anaerobic biofilms grown under conditions of the cystic fibrosis lung. Physiological relevance of ORF PA3911 was demonstrated, inter alia, using phenotype microarray experiments. The mutant strain showed increased susceptibility in the presence of antimicrobials (minocycline, nafcillin, oxacillin, chloramphenicol and thiamphenicol), enhanced twitching motility and significantly impaired biofilm formation. PA3911 is a soluble, cytoplasmic protein in P. aeruginosa In protein-lipid overlay experiments, purified PA3911 bound specifically to phosphatidic acid (PA), the central hub of phospholipid metabolism. Structure-guided site-directed mutagenesis was used to explore the proposed ligand-binding cavity of PA3911. Protein variants of Leu56, Leu58, Val69 and Leu114 were shown to impair PA interaction. A comparative shotgun lipidomics approach demonstrated a multifaceted response of P. aeruginosa to anaerobic conditions at the lipid head group and fatty acid level. Lipid homeostasis in the PA3911 mutant strain was imbalanced with respect to lysophosphatidylcholine, phosphatidylcholine and diacylglycerol under anaerobic and/or aerobic conditions. The impact of the newly identified PA-binding protein on lipid homeostasis and the related macroscopic phenotypes of P. aeruginosa are discussed.

KEYWORDS:

Pseudomonas aeruginosa; lipid-binding protein; lipidomics; site-directed mutagenesis

PMID:
29717024
DOI:
10.1042/BCJ20180257

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