Figure 5.

Recombinant HsPex14p is recognized specifically by ab-MF3 and binds HsPex5p in a ligand blot. Equal amounts of bacterial lysates expressing no recombinant protein (1), biotinylated HsPex14p (2), the biotinylated PTS1-binding domain of HsPex5p (3), or the biotinylated SH3 domain of HsPex13p (4) were subjected to SDS/PAGE and transferred to nitrocellulose. These membranes were then probed with ab-MF3 (A), preimmune serum (B), streptavidin-alkaline phosphatase (C), or [³⁵S]Pex5p in the absence (D) or presence (E) of 60 μM CRYHLKPLQSKL, and reactive bands were visualized as described under Materials and Methods. The migration of molecular mass markers (in kDa) is indicated. The arrow indicates the recombinant expressed, biotinylated HsPex14p; the dot indicates the migration of a nonspecific, immunoreactive bacterial protein (A and B).