title: Inhibition by melittin of phosphorylation by protein kinase C of annexin I from cow mammary gland.
abstract: Protein kinase C (PKC) is a phosphotransferase activated by diacylglycerols, phospholipids and Ca(2+), that regulates a wide variety of biological functions by phosphorylating multiple protein substrates such as annexin I. Annexin I is a phospholipid/Ca (2+)-binding protein distributed in various tissues, including the mammary gland, and is thought to mediate the anti-inflammatory actions of glucocorticoids by inhibiting phospholipase A(2). Melittin, a phospholipase A(2) activator in bee venom, is known to inhibit PKC activity when lysine-rich histone is used as the substrate. The purpose of the present study was to examine whether phosphorylation by PKC of annexin I from cow mammary gland was inhibited by melittin. Melittin inhibited annexin I phosphorylation by PKC in a dose-dependent manner, and its IC(50) value (concentration causing 50% inhibition) was 0.8 microM. The phosphorylation of annexin I was also inhibited by the amphiphilic polypeptides mastoparan and polymyxin B, and their inhibitory effects were comparable to that of melittin. The surface-inactive polypeptide bacitracin was less effective. The inhibition by melittin was effectively reversed by the excess addition of phosphatidylserine, but not distinctly by 1-oleoyl-2-acetyl-sn-glycerol or Ca(2+), suggesting that melittin inhibited the phosphorylation of annexin I by interacting with phosphatidylserine. The inhibition by melittin of PKC phosphorylation of annexin I seems to be pathophysiologically important, because a melittin-like phospholipase A(2)-stimulatory protein is present in bovine endothelial cells.
authors.affiliation: Hokkaido Research Station, National Institute of Animal Health, Hitsujigaoka, Sapporo, Japan.
authors.forename: Norio
authors.initials: N
authors.lastname: Katoh
mesh_terms: D000818:Animals; D017305:Annexin A1 / Q000378:metabolism*; D002417:Cattle; D005260:Female; D020128:Inhibitory Concentration 50; D008321:Mammary Glands, Animal / Q000737:chemistry*; D008555:Melitten / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000378:metabolism*
chemical_list: D017305:Annexin A1; D008555:Melitten; D011493:Protein Kinase C