Rat heart lipoprotein lipase.
Rat heart lipoprotein lipase was highly purified by affinity chromatography using heparin-Sepharose 4B. When extracts of acetone powder were applied to columns, lipase activity was firmly bound to the gel matrix and was later eluted with 1.5 M NaCl. At this stage the eluted enzyme was purified 1500-fold. Disc gel electrophoresis yielded a single protein band corresponding with the enzyme activity. The apparent molecular weight was 60,000. The purified enzyme was highly unstable; however, its activity could be partially stabilized by glycerol or ethylene glycol. In studying the purified enzyme we observed: (i) a cofactor in serum was required for full enzymatic activity; ApoLp-Glu could be substituted for this cofactor; (ii) ApoLp-Ser was inhibitory to lipase activity; (iii) NaCl and protamine sulfate also markedly inhibited the lipase activity; (iv) heparin stimulated the enzymatic activity.
D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008071:Lipoprotein Lipase / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D008970:Molecular Weight; D009206:Myocardium / Q000201:enzymology*; D051381:Rats; D014280:Triglycerides / Q000378:metabolism
D004791:Enzyme Inhibitors; D014280:Triglycerides; D008071:Lipoprotein Lipase