Monoclonal antibodies against rat platelet phospholipase A2.
Monoclonal antibodies which bind specifically to rat platelet phospholipase A2 have been raised. None of them bound to exocrine phospholipase A2 derived from pancreas or snake venom. All antibodies recognized the conformational structure of rat platelet phospholipase A2 supported by intramolecular disulfide bonds, since the reactivity between the antibodies and the enzyme was lost in the presence of 2-mercaptoethanol. One of them, designed MB5.2, inhibited the activity of the platelet phospholipase A2 in a dose-dependent manner. A kinetic study revealed that antibody MB5.2 apparently competed with the substrate for the active site of the enzyme. The other antibodies, designed MD7.1 and ME6.1, inhibited the binding of the enzyme to heparin. The distribution of phospholipases A2 bearing a similar determinant to rat platelet phospholipase A2 was investigated by immunoprecipitation of the enzyme activity or by an immunoblot technique. Among rat tissues, cross-reactivity was observed with phospholipases A2 from spleen, lung, and bone marrow. Extracellular phospholipase A2 detected in the peritoneal cavity of casein-treated rat was also recognized by these antibodies. Furthermore, antibody MD7.1 cross-reacted with rabbit and guinea pig platelet phospholipases A2.
Murakami
M
M
Department of Health Chemistry, Faculty of Pharmaceutical Sciences, University of Tokyo.
Kobayashi
T
T
Umeda
M
M
Kudo
I
I
Inoue
K
K
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001792:Blood Platelets / Q000201:enzymology*; D003429:Cross Reactions; D005260:Female; D006168:Guinea Pigs; D008297:Male; D051379:Mice; D009928:Organ Specificity; D010740:Phospholipases / Q000276:immunology*; D010741:Phospholipases A / Q000097:blood / Q000276:immunology*; D054467:Phospholipases A2; D011817:Rabbits; D051381:Rats; D012685:Sepharose / Q000031:analogs & derivatives
D000911:Antibodies, Monoclonal; C037405:heparin-sepharose; D012685:Sepharose; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2