Characteristics and possible functions of mast cell phospholipases A2.
Phospholipase A2 activity in lysates of mast cells and their related cells [mouse bone marrow-derived IL-3 dependent mast cells (BMMC), rat connective tissue mast cells (CTMC), and rat mastocytoma RBL-2H3 cells] was measured using phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylcholine (PC) as exogenous substrates. Both BMMC and RBL cells showed rather high phospholipase A2 activity, whereas CTMC showed only weak activity. These cells contained at least three types of phospholipase A2. Type 1 enzyme showed no appreciable affinity to heparin, and preferentially hydrolyzed either PC or PE, both of which have an arachidonic acid at the sn-2 position. The activity was absorbed by monoclonal antibody against rabbit platelet cytosolic 85-kDa phospholipase A2. Type 2 enzyme had an affinity to heparin, and was completely inhibited by anti-rat platelet 14-kDa secretory phospholipase A2. This enzyme could be expressed as an "ecto-type" enzyme on the cell surface and might be secreted from cells when mast cells are activated. Type 3 enzyme also had an affinity to heparin, but was separated from type 2 enzyme on reverse-phase HPLC. This enzyme did not interact with anti-14-kDa secretory enzyme antibody. Purified type 3 enzyme (30-kDa) specifically hydrolyzed PS. p-Bromophenacylbromide inhibited all types of phospholipase A2, whereas mepacrine inhibited type 2 and type 3 enzymes, but not type 1 enzyme. Type 2 enzyme was also inhibited by the specific antibody, complement degradation product, and a small-molecular-weight inhibitor. Histamine release was inhibited by all these inhibitors, whereas PGD2 production was inhibited only by p-bromophenacylbromide. Possible roles for these phospholipases A2 in mast cell function are proposed.
Murakami
M
M
Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Kudo
I
I
Inoue
K
K
D000818:Animals; D001853:Bone Marrow / Q000201:enzymology; D001854:Bone Marrow Cells; D002462:Cell Membrane / Q000201:enzymology; D003238:Connective Tissue / Q000201:enzymology; D003239:Connective Tissue Cells; D008407:Mast Cells / Q000201:enzymology*; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000145:classification / Q000378:metabolism*; D054467:Phospholipases A2; D014407:Tumor Cells, Cultured / Q000201:enzymology
D010741:Phospholipases A; D054467:Phospholipases A2