A micromethod for assay of lipoprotein lipase activity in needle biopsy samples of human adipose tissue and skeletal muscle.
A rapid and simple procedure for assay of lipoprotein lipase (LPL) activity in small amounts of human adipose tissue and skeletal muscle is described and validated. The enzyme is eluted from tissues with heparin and the activity is determined from the eluate by measuring the release of [14C]oleic acid from a gum arabic stabilized emulsion of glycerol-tri[14C]oleate in a Tris-buffer medium containing albumin and pooled normal human serum. Reproducible results are obtained with amounts of tissue ranging from 2 to 25 mg. The Km values of the adipose tissue and skeletal muscle LPL for the triolein substrate were 0.74 +/- 0.06 and 0.77 +/- 0.05 mmol/l, respectively. The standard radioactive triolein emulsion was hydrolyzed by adipose tissue LPL at a rate closely similar to rat VLDL-triglyceride labeled in vivo with [1-14C]palmitic acid, suggesting that the experimental substrate behaved in a similar manner to the natural substrate. The LPL activity was much higher in adipose tissue than in muscle. In adipose tissue the LPL activity was 2--4 times higher in women than in men whereas no sex difference was present in the LPL activity of muscle.
D000273:Adipose Tissue / Q000201:enzymology*; D000328:Adult; D001707:Biopsy, Needle; D005260:Female; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D008071:Lipoprotein Lipase / Q000032:analysis*; D008297:Male; D008722:Methods; D008875:Middle Aged; D009132:Muscles / Q000201:enzymology*
D006493:Heparin; D008071:Lipoprotein Lipase