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, deli vering free fatty acids for their use as energy substrates. HSL activi ty is under acute hormonal and neural control, mediated through revers ible phosphorylation of the enzyme. Emerging data from clinical studie s indicate that HSL deficiency or malfunction is associated with sever al pathological situations in humans. In order to perform a biochemica l characterization of human HSL, and to elucidate its molecular proper ties, 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 valuabl e tool for a thorough molecular characterization of the enzyme. (C) 19 98 Academic Press.