Purification and characterization of membrane-bound phospholipase A2 from rat platelets.
Phospholipase A2 was solubilized from rat platelet membrane by 1 M KCl and purified to near homogeneity on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and HPLC. The characteristics of the purified membrane-bound enzyme were compared with those of phospholipase A2 released from thrombin-stimulated rat platelets (Horigome, K., Hayakawa, M., Inoue, K., & Nojima, S. (1987) J. Biochem. 101, 625-631). The molecular weights, elution profiles on reversed-phase HPLC, and NH2-terminal sequences were identical for the two enzymes. Other characteristics of the two enzymes, such as specific activity, substrate specificity, pH optimum, Ca2+ requirement, heat lability, and sensitivity to p-bromophenacyl bromide were also indistinguishable. These findings suggest that both enzymes share a common structure.
Hayakawa
M
M
Department of Health Chemistry, Faculty of Pharmaceutical Sciences, University of Tokyo.
Kudo
I
I
Tomita
M
M
Inoue
K
K
D000818:Animals; D001792:Blood Platelets / Q000201:enzymology*; D002462:Cell Membrane / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D008970:Molecular Weight; D010740:Phospholipases / Q000302:isolation & purification*; D010741:Phospholipases A / Q000097:blood / Q000302:isolation & purification*; D054467:Phospholipases A2; D051381:Rats; D011919:Rats, Inbred Strains; D013010:Sonication; D013917:Thrombin / Q000494:pharmacology
D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D013917:Thrombin