Purification and characterization of a cytosolic, 42-kDa and Ca2+-dependent phospholipase A2 from bovine red blood cells: its involvement in Ca2+-dependent release of arachidonic acid from mammalian red blood cells

The Journal of Biological Chemistry
Hae Sook ShinDae Kyong Kim

Abstract

It has become evident that a Ca(2+)-dependent release of arachidonic acid (AA) and subsequent formation of bioactive lipid mediators such as prostaglandins and leukotrienes in red blood cells (RBCs) can modify physiological functions of neighboring RBCs and platelets. Here we identified a novel type of cytosolic PLA(2) in bovine and human RBCs and purified it to apparent homogeneity with a 14,000-fold purification. The purified enzyme, termed rPLA(2), has a molecular mass of 42 kDa and reveals biochemical properties similar to group IV cPLA(2), but shows different profiles from cPLA(2) in several column chromatographies. Moreover, rPLA(2) did not react with any of anti-cPLA(2) and anti-sPLA(2) antibodies and was identified as an unknown protein in matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis. Divalent metal ions tested exhibited similar effects between rPLA(2) and cPLA(2), whereas mercurials inhibited cPLA(2) but had no effect on rPLA(2). Antibody against the 42-kDa protein not only precipitated the rPLA(2) activity, but also reacted with the 42-kDa protein from bovine and human RBCs in immunoblot analysis. The 42-kDa protein band was selectively detected in murine fetal liver cells kno...Continue Reading

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Citations

Nov 1, 2003·Arteriosclerosis, Thrombosis, and Vascular Biology·Francesco CipolloneCarlo Patrono
Jun 15, 2007·British Journal of Pharmacology·H JiangJ C McGiff
Jul 16, 2010·Pharmacological Reports : PR·Houli JiangJohn C McGiff
Mar 3, 2005·Journal of Cell Science·Philipp A LangThomas Wieder
Dec 14, 2006·Prostaglandins & Other Lipid Mediators·Houli Jiang

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