Table 1. Kinectics and affinity analysis of HC/DC and the indicated mutants to ND-Syt or free Syt II (1–61).
| Ligand | Analyte | KD (M) | ka (1/Ms) | ka Error | kd (1/s) | kd Error |
|---|---|---|---|---|---|---|
| ND-Syt | HC/DC WT | 4.7E−06 | 2.5E+04 | 1.7E+03 | 1.2E−01 | 2.7E−03 |
| HC/DC Y1251A | 3.3E−05 | 9.1E+03 | 4.0E+03 | 3.0E−01 | 1.7E−02 | |
| HC/DC W1252A | 4.8E−05 | 3.2E+03 | 3.7E+03 | 1.6E−01 | 1.4E−02 | |
| HC/DC F1253A | 1.9E−05 | 1.4E+04 | 2.2E+03 | 2.6E−01 | 8.9E−03 | |
| HC/DC F1253W | 4.5E−06 | 2.7E+04 | 1.4E+03 | 1.2E−01 | 1.6E−03 | |
| Syt II (1–61) | HC/DC WT | 4.8E−05 | 6.4E+03 | 1.0E+03 | 3.1E−01 | 1.3E−02 |
| HC/DC Y1251A | 7.5E−05 | 5.5E+03 | 1.7E+03 | 4.1E−01 | 2.7E−02 | |
| HC/DC W1252A | 2.6E−05 | 1.5E+04 | 5.3E+03 | 4.0E−01 | 4.0E−02 | |
| HC/DC F1253A | 2.7E−05 | 1.3E+04 | 2.1E+03 | 3.5E−01 | 1.9E−02 | |
| HC/DC F1253W | 3.7E−05 | 8.0E+03 | 9.6E+02 | 3.0E−01 | 1.1E−02 |
Abbreviations: HC, C-terminal receptor-binding domain; ka, association rate constant; kd, dissociation rate constant; KD, equilibrium dissociation constant; ND, nanodisc; Syt, synaptotagmin; WT, wild-type