TABLE 2.
Predicted binding potential and qualitative evaluation of binding of point-mutated MBP-TbpB to HRP-htf
| Site(s) of mutation(s) | No. of RBD | Predicted impact of mutation on binding potential of RBD | Relative binding of point-mutated TbpB to htfa |
|---|---|---|---|
| Wild type | 100 | ||
| K51N | 1 | Reduction | 104 |
| R54N | 1 | Suppression | 46 |
| R55N | 1 | Suppression | 0 |
| K76N/K77N | 2 | Reduction | 87 |
| R80N | 2 | Suppression | 125 |
| R240N | 3 | Reduction | 42 |
| R243N | 3 | Suppression | 0 |
| MBP (control) | 0 |
a
Crude extracts of equivalent amounts of wild-type and mutant MBP-TbpB were analyzed by Western blotting, probed with HRP-htf (Fig. 3), and quantified by densitometric analysis; the htf binding is expressed as a percentage of the wild-type MBP-TbpB, which is considered 100% binding.