Mapping the lipolytic proteome of adipose tissue using fluorescent suicide inhibitors.
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.
Schicher
Maximilian
M
Institute of Biochemistry, University of Graz, Graz, Austria.
Kollroser
Manfred
M
Hermetter
Albin
A
D000273:Adipose Tissue / Q000737:chemistry* / Q000201:enzymology; D000478:Alkylation; D000818:Animals; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000737:chemistry*; D007525:Isoelectric Focusing; D008049:Lipase / Q000378:metabolism; D008055:Lipids / Q000737:chemistry*; D051379:Mice; D010084:Oxidation-Reduction; D020543:Proteome; D053719:Tandem Mass Spectrometry
D005456:Fluorescent Dyes; D008055:Lipids; D020543:Proteome; D008049:Lipase