Interaction of phospholipase A of the E. coli outer membrane with the inhibitors of eucaryotic phospholipases A₂ and their effect on the Ca²⁺-induced permeabilization of the bacterial membrane.
Phospholipase A of the bacterial outer membrane (OMPLA) is a β-barrel membrane protein which is activated under various stress conditions. The current study examines interaction of inhibitors of eucaryotic phospholipases A₂--palmitoyl trifluoromethyl ketone (PACOCF₃) and aristolochic acid (AA)--with OMPLA and considers a possible involvement of the enzyme in the Ca²⁺-dependent permeabilization of the outer membrane of Escherichia coli. Using the method of molecular docking, it has been predicted that PACOCF₃ and AA bind to OMPLA at the same site and with the same affinity as the OMPLA inhibitors, hexadecanesulfonylfluoride and bromophenacyl bromide, and the substrate of the enzyme palmitoyl oleoyl phosphatidylethanolamine. It has also been shown that PACOCF₃, AA, and bromophenacyl bromide inhibit the Ca²⁺-induced temperature-dependent changes in the permeability of the bacterial membrane for the fluorescent probe propidium iodide and suppressed the transformation of E. coli cells with plasmid DNA induced by Ca²⁺ and heat shock. The cell viability was not affected by the eucaryotic phospholipases A₂ inhibitors. The study discusses a possible involvement of OMPLA in the mechanisms of bacterial transmembrane transport based on the permeabilization of the bacterial outer membrane.
Institute of Theoretical and Experimental Biophysics RAS, Institutskaya, 3, Pushchino, Moscow Region, 142290, Russia, bekonik@gmail.com.
Konstantin N
KN
Belosludtsev
Natalia V
NV
Belosludtseva
Maxim S
MS
Kondratyev
Alexey V
AV
Agafonov
Yuriy A
YA
Purtov
D034341:Aristolochic Acids / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002463:Cell Membrane Permeability / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D007659:Ketones / Q000494:pharmacology; D008958:Models, Molecular; D008968:Molecular Conformation; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D054467:Phospholipases A2 / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D014169:Transformation, Bacterial / Q000187:drug effects
D034341:Aristolochic Acids; D004791:Enzyme Inhibitors; D007659:Ketones; C092652:aristolochic acid C; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium