Table 3.
Pex5p mutants are selectively disturbed in two-hybrid interaction with Pex13-SH3
| GAL4 AD fusion | GAL4 DB fusion | β-Galactosidase activity (RLU/mg protein ± SD) | Growth on his− |
|---|---|---|---|
| Pex5p(F208L) | — | 1.2 ± 0.1 | - |
| Pex5p | Pex13-SH3 | 4,800 ± 800 | + |
| Pex5p(T109A) | Pex13-SH3 | 3,300 ± 300 | + |
| Pex5p(E212V) | Pex13-SH3 | 2.0 ± 0.1 | +/− |
| Pex5p(F208L) | Pex13-SH3 | 1.7 ± 0.1 | - |
| Pex5p(W204A) | Pex13-SH3 | 2.3 ± 0.5 | +/- |
| Pex5p | Mdh3p | 6,900 ± 2,600 | + |
| Pex5p(F208L) | Mdh3p | 5,100 ± 500 | + |
| Pex5p | Pex14p | 10,300 ± 400 | + |
| Pex5p(F208L) | Pex14p | 3,200 ± 400 | + |
| Pex5p(W204A) | Pex14p | 12,000 ± 900 | ND |
Plasmids encoding GAL4 DB and GAL4 AD fusions as indicated were transformed to yeast two-hybrid strains PCY2 and HF7c. Two-hybrid interaction was quantitated in PCY2 by measuring β-galactosidase activity. Indicated is the mean of two measurements of duplo cultures ± SD. Interaction was also measured in HF7c by growth in the absence of histidine (see legend to Table 2). RLU, relative light units.