Positional specificity of hormone-sensitive lipase from rat adipose tissue.

Hormone-sensitive lipase, purified from rat adipose tissue (Fredrikson, G., Strålfors, P., Nilsson, N. O., and Belfrage, P. (1981) J. Biol. Chem. 256, 6311-6320), has been incubated with tri-, di-, and monooleoyl[3H]glycerol, and the acylglycerol reaction products were isolated by thin layer chromatography on silicic acid, impregnated with boric acid. Trioleoylglycerol was hydrolyzed with the intermediate accumulation of monooleoylglycerol, mainly the 2-isomer, and a small amount of 1,2(2,3)-, but no measurable, 1,3-dioleoylglycerol. 2-Monooleoylglycerol was also the major acylglycerol reaction product from 1,2(2,3)-dioleoylglycerol hydrolysis, which occurred at a Vmax of 60% of that with the 1,3-isomer. Part of the 1(3)-monooleoylglycerols found were formed by acyl migration, but 2-ester bond cleavage was directly demonstrated by the use of 1,3-dioleoyl-2-[14C]oleoyl[3H]glycerol as substrate, and by determination of the 14C/3H ratios of the acylglycerol reaction products. Based on the hydrolysis of specific monooleoylglycerol isomers, it was estimated that the 1(3)-ester bonds of the acylglycerols were hydrolyzed 3- to 4-fold faster than the 2-ester bonds. The main lipolytic reaction sequence catalyzed by hormone-sensitive lipase is thus triacylglycerol leads to 1,2(2,3)-diacylglycerol leads to 2-monoacylglycerol. However, the preference for the 1(3)-ester bonds is less marked than that of, e.g. pancreatic and lipoprotein lipase.