Purification of rat liver mitochondrial phospholipase A2.
Rat liver mitochondrial phospholipase A2 was purified to near homogeneity by a combination of gel-filtration, hydroxyapatite and Matrex gel Blue A column chromatography. The absolute positional specificity of the enzyme for acylester bonds at the sn-2-position was established in experiments with 1-[9,10-3H2]palmitoyl-2-[1-14C]linoleoylphosphatidylethanolamine. Molecular weight estimations revealed Mr values of 15000 by SDS-polyacrylamide gel electrophoresis and of 9700 gel by gel-filtration over Ultrogel AcA 54 columns. The enzyme is unaffected by diisopropylfluorophosphate and thiol reagents such as 5,5'-dithiobis(2-nitrobenzoic acid), N-ethylmaleimide and iodoacetamide, but is completely inhibited by the alkylating reagent p-bromophenacylbromide.
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002850:Chromatography, Gel; D006358:Hot Temperature; D007700:Kinetics; D008930:Mitochondria, Liver / Q000201:enzymology*; D008970:Molecular Weight; D010740:Phospholipases / Q000302:isolation & purification*; D010741:Phospholipases A / Q000302:isolation & purification*; D054467:Phospholipases A2; D051381:Rats; D012995:Solubility; D013329:Structure-Activity Relationship
D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium