TY  - JOUR
AU  - Bleffert, Florian
AU  - Granzin, Joachim
AU  - Caliskan, Muttalip
AU  - Schott-Verdugo, Stephan N
AU  - Siebers, Meike
AU  - Thiele, Björn
AU  - Rahme, Laurence G
AU  - Felgner, Sebastian
AU  - Dörmann, Peter
AU  - Gohlke, Holger
AU  - Batra-Safferling, Renu
AU  - Jaeger, Karl-Erich
AU  - Kovacic, Filip
TI  - Structural, mechanistic and physiological insights into phospholipase A-mediated membrane phospholipid degradation in Pseudomonas aeruginosa
JO  - eLife
VL  - 11
SN  - 2050-084X
CY  - Cambridge
PB  - eLife Sciences Publications
M1  - FZJ-2022-02211
SP  - e72824
PY  - 2022
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:35536643
UR  - <Go to ISI:>//WOS:000804454400001
DO  - DOI:10.7554/eLife.72824
UR  - https://juser.fz-juelich.de/record/907782
ER  -