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. 2012 May 1;22(7):1155–1168. doi: 10.1038/cr.2012.71

Figure 5.

Figure 5

CBL2 is modified by stearate and palmitate. CBL2-GFP and a mutated version of CBL2 harboring the triple cysteine to serine exchanges (C4,12,18S) were transiently expressed in N. benthamiana leaves. Native proteins were extracted from the leaf material and GFP-tagged proteins were enriched by differential ammonium sulfate precipitations and ion-exchange chromatography. Lipid groups were removed by hydrogenation and analyzed by GC coupled MS analyses. (A) GC retention and identification of standard ethyl palmitate (after 32 min) and ethyl stearate (after 34.3 min) derivatives (bottom chromatogram), and of the hydrogenated lipids removed from CBL2-GFP and CBL2C4,12,18S-GFP. (B) MS chromatograms of the ethyl palmitate and ethyl stearate standards (top panels) and of ethyl palmitate and ethyl stearate released from CBL2-GFP (bottom panels).