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. Author manuscript; available in PMC: 2011 Apr 6.
Published in final edited form as: Biochemistry. 2010 Apr 6;49(13):2925–2931. doi: 10.1021/bi9021439

Figure 5. Bioinylation of PCFT mutants with MTSEA-biotin after permeabilization.

Figure 5

Panel A: HeLa cells transiently expressing CL-PCFT or various mutants were permeabilized with digitonin before the biotinylation reaction. Panel B: Dependence of biotinylation inhibition by MTSES on digitonin permeabilization. Three conditions of biotinylation labeling were conducted for each mutant: (1) Cells were permeabilized before the biotinylation reaction (same as described for panel A of this figure). (2) Cells were first treated with MTSES, then permeabilized with digitinon before the biotinylation reaction. (3) Cells were permeabilized with digitonin first, then treated with MTSES before the biotinylation reaction. For all panels, the crude membrane sample was matched to the precipitated sample and results are representative of at least two separate experiments.