A phospholipase B from Pseudomonas aeruginosa with activity towards endogenous phospholipids affects biofilm assembly

Weiler, Andrea Jeanette; Spitz, Olivia; Gudzuhn, Mirja; Schott‐Verdugo, Stephan; Kamel, Michael; Thiele, Björn; Streit, Wolfgang R.; Kedrov, Alexej; Schmitt, Lutz; Gohlke, Holger; Kovačić, Filip

doi:10.1016/j.bbalip.2021.159101

Cited by 9 publications

(9 citation statements)

References 78 publications

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“…Although PlaF is an important enzyme involved in GPL metabolism, future research should reveal (i) which acyltransferase is involved in the acylation of lysoGPLs produced by PlaF, (ii) if PlaF has acyltransferase activity as described for cPLA 2 γ involved in the Lands cycle in humans ( Asai et al, 2003 ), and (iii) if periplasmic lysophospholipase TesA ( Kovačić et al, 2013 ) and the recently discovered intracellular PLA PlaB ( Weiler et al, 2022 ) are involved in the Lands cycle.…”

Section: Discussionmentioning

confidence: 99%

“…However, this enzyme has no PLA activity, and therefore it is most likely not related to membrane GPL remodeling ( Leščić Ašler et al, 2010 ). We recently published a novel intracellular PLA from P. aeruginosa whose function for remodeling of GPLs still needs to be experimentally analyzed ( Weiler et al, 2022 ). Comprehensive lipidomic profiling of 113 Escherichia coli strains with deleted or overexpressed lipid metabolism genes did not reveal the identity of an intracellular PLA suitable for the Lands cycle ( Jeucken et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

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Structural, mechanistic, and physiological insights into phospholipase A-mediated membrane phospholipid degradation in Pseudomonas aeruginosa

Bleffert

Granzin

Caliskan

et al. 2022

eLife

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132

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Cells steadily adapt their membrane glycerophospholipid (GPL) composition to changing environmental and developmental conditions. While the regulation of membrane homeostasis via GPL synthesis in bacteria has been studied in detail, the mechanisms underlying the controlled degradation of endogenous GPLs remain unknown. Thus far, the function of intracellular phospholipases A (PLAs) in GPL remodeling (Lands cycle) in bacteria is not clearly established. Here, we identified the first cytoplasmic membrane-bound phospholipase A1 (PlaF) from Pseudomonas aeruginosa, which might be involved in the Lands cycle. PlaF is an important virulence factor, as the P. aeruginosa ΔplaF mutant showed strongly attenuated virulence in Galleria mellonella and macrophages. We present a 2.0-Å-resolution crystal structure of PlaF, the first structure that reveals homodimerization of a single-pass transmembrane (TM) full-length protein. PlaF dimerization, mediated solely through the intermolecular interactions of TM and juxtamembrane regions, inhibits its activity. The dimerization site and the catalytic sites are linked by an intricate ligand-mediated interaction network, which might explain the product (fatty acid) feedback inhibition observed with the purified PlaF protein. We used molecular dynamics simulations and configurational free energy computations to suggest a model of PlaF activation through a coupled monomerization and tilting of the monomer in the membrane, which constrains the active site cavity into contact with the GPL substrates. Thus, these data show the importance of the PlaF mediated GPL remodeling pathway for virulence and could pave the way for the development of novel therapeutics targeting PlaF.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Structural, mechanistic, and physiological insights into phospholipase A-mediated membrane phospholipid degradation in Pseudomonas aeruginosa

Bleffert

Granzin

Caliskan

et al. 2022

eLife

Self Cite

132

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“…MALS was performed as per the previous report [55]. Superdex 200 Increase 10/300 GL column (GE Healthcare) was pre-equilibrated overnight at 0.1 ml/minute flow rate with buffer E (20 mM HEPES pH 8.0, 200 mM NaCl, 1 mM β-ME).…”

Section: Multiangle Light Scattering (Mals)mentioning

confidence: 99%

A MademoiseLLE domain binding platform links the key RNA transporter to endosomes

et al. 2022

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Spatiotemporal expression can be achieved by transport and translation of mRNAs at defined subcellular sites. An emerging mechanism mediating mRNA trafficking is microtubule-dependent co-transport on shuttling endosomes. Although progress has been made in identifying various components of the endosomal mRNA transport machinery, a mechanistic understanding of how these RNA-binding proteins are connected to endosomes is still lacking. Here, we demonstrate that a flexible MademoiseLLE (MLLE) domain platform within RNA-binding protein Rrm4 of Ustilago maydis is crucial for endosomal attachment. Our structure/function analysis uncovered three MLLE domains at the C-terminus of Rrm4 with a functionally defined hierarchy. MLLE3 recognises two PAM2-like sequences of the adaptor protein Upa1 and is essential for endosomal shuttling of Rrm4. MLLE1 and MLLE2 are most likely accessory domains exhibiting a variable binding mode for interaction with currently unknown partners. Thus, endosomal attachment of the mRNA transporter is orchestrated by a sophisticated MLLE domain binding platform.

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“…Here, we show that the PldA of C. jejuni possesses phospholipase activity that generates ( Sn )-1 and ( Sn) -2 LPLs which is more typical for phospholipase class B proteins. 20 We also for the first time provide evidence that the produced lysoPE phospholipids can lyse erythrocytes and damage epithelial cells. This effect is especially evident for short-chain lysoPE species and can be rescued by oxidative stress inhibitors.…”

Section: Discussionmentioning

confidence: 60%

Campylobacter jejuni permeabilizes the host cell membrane by short chain lysophosphatidylethanolamines

et al. 2022

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Lysophospholipids (LPLs) are crucial for regulating epithelial integrity and homeostasis in eukaryotes, however the effects of LPLs produced by bacteria on host cells is largely unknown. The membrane of the human bacterial pathogen Campylobacter jejuni is rich in LPLs. Although C. jejuni possesses several virulence factors, it lacks traditional virulence factors like type III secretion systems, present in most enteropathogens. Here, we provide evidence that membrane lipids lysophosphatidylethanolamines (lysoPEs) of C. jejuni are able to lyse erythrocytes and are toxic for HeLa and Caco-2 cells. Lactate dehydrogenase (LDH) release assays and confocal microscopy revealed that lysoPE permeabilizes the cells. LysoPE toxicity was partially rescued by oxidative stress inhibitors, indicating that intracellular reactive oxygen species may contribute to the cell damage. Our results show that especially the short-chain lysoPEs (C:14) which is abundantly present in the C. jejuni membrane may be considered as a novel virulence factor.

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A phospholipase B from Pseudomonas aeruginosa with activity towards endogenous phospholipids affects biofilm assembly

Cited by 9 publications

References 78 publications

Structural, mechanistic, and physiological insights into phospholipase A-mediated membrane phospholipid degradation in Pseudomonas aeruginosa

Structural, mechanistic, and physiological insights into phospholipase A-mediated membrane phospholipid degradation in Pseudomonas aeruginosa

A MademoiseLLE domain binding platform links the key RNA transporter to endosomes

Campylobacter jejuni permeabilizes the host cell membrane by short chain lysophosphatidylethanolamines

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