Fig 4.

Recombinant CrLIP1 degrades diacylglycerol in vitro. (A) Thin-layer chromatograph of lipids extracted from diacylglycerol lipase assay mixtures with diolein as the substrate for CrLIP1. Lane 1, control reaction with diolein and CrLIP1 storage buffer; lane 2, reaction with diolein and purified CrLIP1 protein; lane 3, control reaction with purified CrLIP1 protein without diolein to show fatty acids contained in the protein preparation. Diolein and oleic acid (18:1Δ⁹) were loaded as markers. The deep staining at the origins might be from ingredients of the reaction buffer, such as dithiothreitol or glycerol. (B to D) Gas-liquid chromatographs of fatty acid methyl esters (FAMEs) derived from free fatty acids scraped from the TLC plate shown in panel A. Arrows point to the location or expected location of oleic FAME.