TABLE 2.
Binding kinetics of GST-NGFpro binding to sortilin mutants
Concentration series of 10, 20, 30, 40, and 50 nm of GST-NGFpro was applied to biosensor chips with immobilized sortilin single residue mutants. Affinities and rate constants were estimated using the BIAevaluation software, using the 1:1 Langmuir binding isotherm.
| Mutation | Amino acids 160–174 | 10 nm to 50 nm series of sensorgrams |
||
|---|---|---|---|---|
| ka | kd | KD | ||
| s−1 | m s−1 | |||
| WT | RGGRIFRSSDFAKNF | 1.72 × 105 | 3.45 × 10−4 | 2.12 nM |
| R160A | AGGRIFRSSDFAKNF | 1.47 × 105 | 2.27 × 10−4 | 1.89 nM |
| R163A | RGGAIFRSSDFAKNF | 1.85 × 105 | 4.21 × 10−4 | 2.70 nM |
| F165A | RGGRIARSSDFAKNF | 1.10 × 105 | 4.22 × 10−4 | 3.86 nM |
| R166A | RGGRIFASSDFAKNF | 6.48 × 104 | 2.85 × 10−4 | 4.38 nM |
| F170A | RGGRIFRSSDAAKNF | 1.30 × 105 | 5.91 × 10−4 | 5.59 nM |
| K172A | RGGRIFRSSDFAANF | 1.01 × 105 | 3.46 × 10−4 | 3.91 nM |
| F174A | RGGRIFRSSDFAKNA | 2.35 × 105 | 1.03 × 10−3 | 4.27 nM |