Mapping the lipolytic proteome of adipose tissue using fluorescent suicide inhibitors

Abstract

Lipases are responsible for the hydrolysis of acylglycerols and cholesteryl esters in animals, plants, and microorganisms. In this chapter we describe a tool for the concomitant analysis of lipases in complex proteomes. For this purpose, the target enzymes are selectively and covalently labelled with fluorescent suicide inhibitors. Stable lipid–protein complexes are formed that are separated by gel electrophoresis and identified by mass spectrometry.