Binding parameters of VASH–SVBP complexes interaction with tyrosinated- and detyrosinated-enriched microtubules in variable ionic strength media
| Experiment (medium) . | Ionic strength (mM) . | Microtubule . | Recombinant protein (concentration) . | Residence time τ (s) . | Binding frequency (min−1.nM−1.µm-1) . |
|---|---|---|---|---|---|
| A (BRB40, 50 mM KCl) | 140 | Tyr-MTs | V1_FL (50 pM) | 0.5 | 80.8 |
| Tyr-MTs | deadV1_FL (50 pM) | 1.0 | 132.2 | ||
| Tyr-MTs | V2_FL (50 pM) | >11 | 13.0 | ||
| Tyr-MTs | deadV2_FL (50 pM) | >14.6 | 13.9 | ||
| B (BRB80) | 173 | Tyr-MTs | V1_FL (50 pM) | 0.8 | 51.6 |
| Tyr-MTs | deadV1_FL (50 pM) | 0.9 | 86.4 | ||
| C (BRB80, 50 mM KCl) | 223 | Tyr-MTs | V1_FL (1 nM) | 0.2 | 3.0 |
| Tyr-MTs | deadV1_FL (1 nM) | 0. 2 | 12.7 | ||
| Tyr-MTs | V2_FL (100 pM) | 3.9 | 20.4 | ||
| Tyr-MTs | deadV2_FL (100 pM) | >7.2 | 13.5 | ||
| D (BRB80, 100 mM KCl) | 273 | Tyr-MTs | V2_FL (50 pM) | 0.9 | 14.9 |
| Tyr-MTs | deadV2_FL (50 pM) | 1.4 | 19.3 | ||
| deTyr-MTs | V2_FL (50 pM) | 0.6 | 13.7 | ||
| deTyr-MTs | deadV2_FL (50 pM) | 0.7 | 41.0 | ||
| E (BRB40, 50 mM KCl) | 140 | Tyr-MTs | V1_FL (50 pM) | 0.8 | 53.4 |
| Tyr-MTs | deadV1_FL (50 pM) | 1.2 | 89.0 | ||
| deTyr-MTs | V1_FL (50 pM) | 0.5 | 42.3 | ||
| deTyr-MTs | deadV1_FL (50 pM) | 0.6 | 66.5 |
| Experiment (medium) . | Ionic strength (mM) . | Microtubule . | Recombinant protein (concentration) . | Residence time τ (s) . | Binding frequency (min−1.nM−1.µm-1) . |
|---|---|---|---|---|---|
| A (BRB40, 50 mM KCl) | 140 | Tyr-MTs | V1_FL (50 pM) | 0.5 | 80.8 |
| Tyr-MTs | deadV1_FL (50 pM) | 1.0 | 132.2 | ||
| Tyr-MTs | V2_FL (50 pM) | >11 | 13.0 | ||
| Tyr-MTs | deadV2_FL (50 pM) | >14.6 | 13.9 | ||
| B (BRB80) | 173 | Tyr-MTs | V1_FL (50 pM) | 0.8 | 51.6 |
| Tyr-MTs | deadV1_FL (50 pM) | 0.9 | 86.4 | ||
| C (BRB80, 50 mM KCl) | 223 | Tyr-MTs | V1_FL (1 nM) | 0.2 | 3.0 |
| Tyr-MTs | deadV1_FL (1 nM) | 0. 2 | 12.7 | ||
| Tyr-MTs | V2_FL (100 pM) | 3.9 | 20.4 | ||
| Tyr-MTs | deadV2_FL (100 pM) | >7.2 | 13.5 | ||
| D (BRB80, 100 mM KCl) | 273 | Tyr-MTs | V2_FL (50 pM) | 0.9 | 14.9 |
| Tyr-MTs | deadV2_FL (50 pM) | 1.4 | 19.3 | ||
| deTyr-MTs | V2_FL (50 pM) | 0.6 | 13.7 | ||
| deTyr-MTs | deadV2_FL (50 pM) | 0.7 | 41.0 | ||
| E (BRB40, 50 mM KCl) | 140 | Tyr-MTs | V1_FL (50 pM) | 0.8 | 53.4 |
| Tyr-MTs | deadV1_FL (50 pM) | 1.2 | 89.0 | ||
| deTyr-MTs | V1_FL (50 pM) | 0.5 | 42.3 | ||
| deTyr-MTs | deadV1_FL (50 pM) | 0.6 | 66.5 |
(A) Related to Fig. 1, D, F, and G. (B) Related to Fig. 2 A. (C) Related to Fig. 2 B. (D) Related to Fig. 2, C, G, and H. (E) related to Fig. 2, E and F. A−E represent experiments performed on different days. At least 15 microtubules were analyzed. See Materials and methods for detailed experimental conditions.