Figure 2

Nucleotide-dependent association of the AAA-complex to Pex15p.

Protein complexes were isolated from 1% digitonin-solubilized membranes of wild-type (control) and wild-type cells expressing Pex15pTPA via IgG-Sepharose and subsequent TEV protease cleavage. Complex isolation was performed in the absence or presence of 5 mM ATP and ADP as indicated. Equal portion of solubilisate (1.2% of total) and eluate (20% of total) were subjected to SDS-PAGE and immunoblot analysis and probed with specific antibodies as indicated. Pex1p and Pex6p were part of the Pex15p complex when ADP or ATP was present, whereas a significant loss of binding to Pex15p was observed in the absence of nucleotides.