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. 2011 Nov 16;6(11):e27815. doi: 10.1371/journal.pone.0027815

Figure 2. Mapping of the Impα isoform domains involved in the interaction with Vpr.

Figure 2

. (A) Schematic representation of the Impα isoforms, Rch1, Qip1, NPI-1 and their deletion mutants. (B and C) All mutants were expressed as GST fusion proteins in E. coli and purified using Glutathione-Sepharose. Twelve pmol of purified GST-tagged Impα isoform derivatives were resolved by 10% SDS-PAGE and stained with CBB. (D and E) Binding of Impα isoforms to Vpr. Glutathione-Sepharose beads coupled to GST-Impα isoforms or GST alone were incubated with mRFP-Vpr or mRFP. The bound fractions and 1/50 of the input of mRFP-Vpr and mRFP were analyzed by immunoblotting with anti-Flag M2 MAb (right panel). The positions of mRFP and mRFP-Vpr are indicated.