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. Author manuscript; available in PMC: 2011 Sep 28.
Published in final edited form as: Biochemistry. 2010 Sep 28;49(38):8338–8349. doi: 10.1021/bi100466q

Figure 2. Physical interaction between Nef and ABCA1 depends upon Nef myristoylation and is disrupted by mutation of sequences between 2225 and 2231 of the ABCA1 C-terminus.

Figure 2

293ET cells were transfected with cDNAs that express wild type FLAG-ABCA1 (FLAG tag located in the N-terminal first extracellular loop of ABCA1) or the indicated FLAG-ABCA1 C-terminal mutants in the presence of the indicated Nef constructs (ABCA1-Δ40: a 40-AA C-terminal deletion, ABCA1-DDDHLK-AAAAAA: sequences between −31 and −36 from the C-terminus selectively mutated to alanines) (NefG2A, alanine mutation of myristoylation motif). FLAG-ABCA1 was immunoprecipitated and the amount of co-precipitated Nef and ABCA1 was determined by immunoblotting (left panels). The amount of ABCA1, Nef and β-actin expressed in the transfected cells was further determined by immunoblotting the input protein lysates (right panels).