Human hormone-sensitive lipase: Expression and large-scale purification from a baculovirus/insect cell system

Hormone-sensitive lipase (HSL) is a key enzyme in lipid metabolism and overall energy homeostasis in mammals. It catalyzes the rate-limiting step in the hydrolysis of triglyceride stores in the adipocytes, delivering free fatty acids for their use as energy substrates. HSL activity is under acute hormonal and neural control, mediated through reversible phosphorylation of the enzyme. Emerging data from clinical studies indicate that HSL deficiency or malfunction is associated with several pathological situations in humans. In order to perform a biochemical characterization of human HSL, and to elucidate its molecular properties, purification of homogeneous protein in large amounts is required. Here, we describe the expression and purification of a catalytically active recombinant human HSL, The process allows the purification of milligram amounts of homogeneous protein, and should provide a valuable tool for a thorough molecular characterization of the enzyme. (C) 1998 Academic Press.