PARTIAL-PURIFICATION AND IDENTIFICATION OF HORMONE-SENSITIVE LIPASE FROM CHICKEN ADIPOSE-TISSUE

HSL from chicken adipose tissue exhibits remarkable activation upon ph osphorylation with cAMP-dependent protein kinase (cAMP-PK) compared La HSL from rat and human adipose tissue. In order to characterize the c hicken HSL enzyme, it was purified 3500 fold from a chicken adipose ti ssue homogenate using pH 5.2 precipitation and anion-exchange chromato graphy. The purified chicken HSL was identified as all 86 kDa protein using Western blot analysis. The HSL diacylglycerol lipase activity wa s inhibited by 98% upon incubation with anti-rat ASE antiserum, and th e specific activity of chicken HSE was estimated to be approximately t he same as for the rat enzyme. Furthermore, the 86 kDa polypeptide was phosphorylated by GAMP-PR to about the same stoichiometry as for the recombinant rat enzyme. Hence, our results demonstrate that HSL from c hicken adipose tissue is comparable in size and specific activity to H SL from mammalian species, and not a smaller 42 kDa polypeptide with 1 000-fold lower specific activity as previously reported (Berglund, L., Khoo, J. C., Jensen, D., and Steinberg, D., 1980 J. Biol. Chem. 255, 5420-5428). (C) 1997 Academic Press.