Lipoprotein lipase from bovine milk. Isolation procedure, chemical characterization, and molecular weight analysis.
Lipoprotein lipase of high purity has been isolated from bovine milk by affinity chromatography on heparin-Sepharose, adsorption to Cgamma-aluminum hydroxide gel, and intervent dilution chromatography on heparin-Sepharose. Chemical analysis shows that the enzyme is a glycoprotein containing 8.3% carbohydrate. The monomer molecular weight, determined under reducing conditions in 6.6 M guanidine HCl by sedimentation equilibrium ultracentrifugation and analytical gel chromatrgraphy, is 48,300 and 50,800, respectively. Analyses of the sedimentation coefficient (SO20,w=5.40 S) and the diffusion coefficient (DO20,w=48.8 mum2/s) in a buffer of physiological pH and ionic strength yield a molecular weight of 96,900. In solution, the native enzyme thus appears to be a dimer of presumably identical subunits.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002417:Cattle; D002846:Chromatography, Affinity; D004355:Drug Stability; D005260:Female; D005990:Glycerol / Q000494:pharmacology; D005998:Glycine / Q000494:pharmacology; D005779:Immunodiffusion; D008071:Lipoprotein Lipase / Q000302:isolation & purification / Q000378:metabolism; D008892:Milk / Q000201:enzymology*; D008970:Molecular Weight
D000596:Amino Acids; D002241:Carbohydrates; D008071:Lipoprotein Lipase; D005990:Glycerol; D005998:Glycine