Two immunologically and catalytically distinct arachidonate 12-lipoxygenases of bovine platelets and leukocytes.
12-Lipoxygenases were found in the cytosol fraction of bovine leukocytes and platelets. The bovine leukocyte enzyme was immunoprecipitable by a monoclonal antibody directed to 12-lipoxygenase of porcine leukocytes, but not by a monoclonal antibody against the human platelet enzyme. In contrast, the bovine platelet enzyme cross-reacted only with antibody against the human platelet enzyme. The leukocyte and platelet enzymes were partially purified to final specific enzyme activities of 1.1 and 0.3 mumol/min/mg protein, respectively, by immunoaffinity chromatography using each cross-reacting antibody as a ligand. The leukocyte enzyme reacted with various octadecapolyenoic acids as well as eicosapolyenoic and docosapolyenoic acids, whereas the platelet enzyme was almost inactive with octadecapolyenoic acids. Moreover, the two enzymes showed different heat-instabilities and reaction time courses. Thus, the 12-lipoxygenases of bovine leukocytes and platelets were immunologically and catalytically distinct enzymes.
Takahashi
Y
Y
Department of Biochemistry, Tokushima University, School of Medicine, Japan.
Ueda
N
N
Yamamoto
S
S
D000818:Animals; D000911:Antibodies, Monoclonal; D001092:Arachidonate 12-Lipoxygenase / Q000097:blood* / Q000145:classification / Q000276:immunology; D001091:Arachidonate Lipoxygenases / Q000097:blood*; D001792:Blood Platelets / Q000201:enzymology*; D002384:Catalysis; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D006358:Hot Temperature; D015151:Immunoblotting; D007962:Leukocytes / Q000201:enzymology*; D013058:Mass Spectrometry; D013056:Spectrophotometry, Ultraviolet
D000911:Antibodies, Monoclonal; D001091:Arachidonate Lipoxygenases; D001092:Arachidonate 12-Lipoxygenase