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. 2009 Feb 13;284(7):4404–4412. doi: 10.1074/jbc.M808645200

FIGURE 2.

FIGURE 2.

PalB binds directly to Vps24. Key residues in the Vps24 MIM domain are essential for PalB binding. A, autoradiographs of GST pull-down assays with the indicated A. nidulans GST fusion proteins and 35S-labeled PalA and PalB preys synthesized in vitro using a coupled transcription/translation system. Input lanes contained 2% of the material used in the pull-downs. Note that GST-DidB does not pull-down PalB under these conditions. B, amino acid sequence comparison of the MIM motifs of fungal Vps24 and Vps2 proteins and their respective CHMP3 and CHMP2A human orthologues is shown on the left. Shaded residues correspond to the consensus MIM motif recognizing the Vps4 MIT domain (38, 42). AnVps24 Arg-222 and Leu-223 correspond to positions 0 and 1 in the consensus motif. C, GST pull-down assays using wild-type or mutant (R222D and L223D substituted) A. nidulans GST::Vps24 fusion proteins as baits and 35S-labeled wild-type PalB as prey.