Unknown

Dataset Information

0

High specificity of human secretory class II phospholipase A2 for phosphatidic acid.


ABSTRACT: Lysophosphatidic acid (LPA) is a potent lipid second messenger which stimulates platelet aggregation, cell proliferation and smooth-muscle contraction. The phospholipase A2 (PLA2)-catalysed hydrolysis of phosphatidic acid (PA) is thought to be a primary synthetic route for LPA. Of the multiple forms of PLA2 present in human tissues, human secretory class-II PLA2 (hs-PLA2) has been implicated in the production of LPA from platelets and whole blood cells challenged with inflammatory stimuli. To explore further the possibility that hs-PLA2 is involved in the production of LPA, we rigorously measured the phospholipid head group specificity of hs-PLA2 by a novel PLA2 kinetic system using polymerized mixed liposomes. Kinetic analysis of recombinant hs-PLA2 demonstrates that hs-PLA2 strongly prefers PA as substrate over other phospholipids found in the mammalian plasma membrane including phosphatidylserine (PS), phosphatidylcholine (PC) and phosphatidylethanolamine (PE). The order of preference is PA >> PE approximately PS > PC. To identify amino acid residues of hs-PLA2 that are involved in its unique substrate specificity, we mutated two residues, Glu-56 and Lys-69, which were shown to interact with the phospholipid head group in the X-ray-crystallographic structure of the hs-PLA2-transition-state-analogue complex. The K69Y mutant showed selective inactivation toward PA whereas the E56K mutant displayed a most pronounced inactivation to PE. Thus it appears that Lys-69 is at least partially involved in the PA specificity of hs-PLA2 and Glu-56 in the distinction between PE and PC. In conjunction with a recent cell study [Fourcade, Simon, Viode, Rugani, Leballe, Ragab, Fournie, Sarda and Chap (1995) Cell 80, 919-927], these studies suggest that hs-PLA2 can rapidly hydrolyse PA molecules exposed to the outer layer of cell-derived microvesicles and thereby produce LPA.

SUBMITTER: Snitko Y 

PROVIDER: S-EPMC1218130 | BioStudies | 1997-01-01

REPOSITORIES: biostudies

Similar Datasets

1995-01-01 | S-EPMC1136425 | BioStudies
2021-01-01 | S-EPMC7851136 | BioStudies
2020-01-01 | S-EPMC7761402 | BioStudies
2011-01-01 | S-EPMC3089575 | BioStudies
2013-01-01 | S-EPMC3770101 | BioStudies
1996-01-01 | S-EPMC1217421 | BioStudies
2001-01-01 | S-EPMC1221700 | BioStudies
2012-01-01 | S-EPMC3277569 | BioStudies
1000-01-01 | S-EPMC2785640 | BioStudies
1993-01-01 | S-EPMC1132423 | BioStudies