Mammalian secretory class V phospholipase A2 (PLA2) is a newly discovered PLA2 that is implicated in eicosanoid formation in inflammatory cells. As a first step towards understanding the structure, function and regulation of this PLA2, we constructed a bacterial expression vector for human secretory class V PLA2 (hV-PLA2), over-expressed and purified the protein, and determined its physical and...

Parfenova, Helena, John Haffner, and Charles W. Leffler. Phosphorylation-dependent stimulation of prostanoid synthesis by nigericin in cerebral endothelial cells. Am. J. Physiol. 277 (Cell Physiol. 46): C728–C738, 1999.— Nigericin decreases intracellular pH (pHi) and stimulates prostanoid (PG) synthesis in endothelial cells from cerebral microvessels of newborn pigs. Nigericin-induced PG produc...

By analyzing human embryonic kidney 293 cell transfectants stably overexpressing various types of phospholipase A2 (PLA2), we have shown that polyunsaturated fatty acids (PUFAs) preferentially activate type IIA secretory PLA2 (sPLA2-IIA)-mediated arachidonic acid (AA) release from interleukin-1 (IL-1)-stimulated cells. When 293 cells prelabeled with 13H]AA were incubated with exogenous PUFAs in...

We determined the in vitro inhibitory effects of cocoa extracts and procyanidins against pancreatic α-amylase (PA), pancreatic lipase (PL) and secreted phospholipase A2 (PLA2), and characterized the kinetics of such inhibition. Lavado, regular and Dutch-processed cocoa extracts as well as cocoa procyanidins (degree of polymerization (DP) = 2 to 10) were examined. Cocoa extracts and procyanidins...

Objective: This research investigates the antibacterial, anticancer, antioxidant, and antiretroviral activities of lionfish spine poison extract.
 Methods: Isolation purification phospholipase A2 (PLA2) protein obtained from were conducted through following stages, including, extraction venom by sonication, heating, using gradual saturation levels ammonium sulfate. Furthermore, purity conc...

Phosphatidylcholine (PtdCho) is a major membrane phospholipid, and its loss is sufficient in itself to induce cell death. PtdCho homeostasis is regulated by the balance between hydrolysis and synthesis. PtdCho is hydrolyzed by phospholipase A2 (PLA2), PtdChospecific phospholipase C (PtdCho-PLC), and phospholipase D (PLD). PtdCho synthesis is rate-limited by CTP:phosphocholine cytidylyltransfera...

The crystal structure of a complex between a phosphonate transition-state analogue and the phospholipase A2 (PLA2) from Naja naja atra venom has been solved and refined to a resolution of 2.0 angstroms. The identical stereochemistry of the two complexes that comprise the crystal's asymmetric unit indicates both the manner in which the transition state is stabilized and how the hydrophobic fatty...

We have developed a simple fluorescent assay for detection of phospholipase A2 (PLA2) activity in zebrafish embryos that utilizes a fluorescent phosphatidylcholine substrate. By using this assay in conjunction with selective PLA2 inhibitors and Western blot analysis, we identified the principal activity in zebrafish embryogenesis as characteristic of the Ca2+-dependent cytosolic PLA2 (cPLA2) su...

We purified an 80-kDa Ca2+-independent phospholipase A2 (iPLA2) from rat brain using octyl-Sepharose, ATP-agarose, and calmodulin-agarose column chromatography steps. This procedure gave a 30,000-fold purification and yielded 4 microg of a near-homogeneous iPLA2 with a specific activity of 4.3 micromol/min/mg. Peptide sequences of the rat brain iPLA2 display considerable homology to sequences o...