Binding parameters on microtubules of full-length, truncated, and chimeric versions of VASH1 and VASH2 enzymes in complex with SVBP
| 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 | deadV1_FL (50 pM) | 1.2 | 80.4 |
| deadV1_Nt + CD (50 pM) | - | No binding | |||
| deadV1_CD (50 pM) | 1.4 | 70.7 | |||
| deadV1_CD + Ct (50 pM) | >7.7 | 21.1 | |||
| deadV2_FL (50 pM) | >17.1 | 16.2 | |||
| deadV2_Nt + CD (50 pM) | >4.0 | 35.9 | |||
| deadV2_CD (50 pM) | 1.2 | 85.1 | |||
| deadV2_CD + Ct (50 pM) | >10.4 | 10.4 | |||
| B (BRB40, 50 mM KCl) | 140 | Tyr-MTs | deadV1_FL (50 pM) | 1.2 | 54.1 |
| deadV1_FL(NtV2) (50 pM) | >9.9 | 31.0 | |||
| deadV2_FL (50 pM) | >13.4 | 19.1 | |||
| deadV2_FL(NtV1) (50 pM) | >6.5 | 18.1 | |||
| C (BRB40, 50 mM KCl) | 140 | Tyr-MTs | V1_FL (25 pM) | 1.2 | 96.7 |
| V1_FL(NtV2) (25 pM) | >10.7 | 20.8 | |||
| V2_FL (25 pM) | >12.9 | 22.9 | |||
| V2_FL(NtV1) (25 pM) | >3.9 | 42.4 |
| 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 | deadV1_FL (50 pM) | 1.2 | 80.4 |
| deadV1_Nt + CD (50 pM) | - | No binding | |||
| deadV1_CD (50 pM) | 1.4 | 70.7 | |||
| deadV1_CD + Ct (50 pM) | >7.7 | 21.1 | |||
| deadV2_FL (50 pM) | >17.1 | 16.2 | |||
| deadV2_Nt + CD (50 pM) | >4.0 | 35.9 | |||
| deadV2_CD (50 pM) | 1.2 | 85.1 | |||
| deadV2_CD + Ct (50 pM) | >10.4 | 10.4 | |||
| B (BRB40, 50 mM KCl) | 140 | Tyr-MTs | deadV1_FL (50 pM) | 1.2 | 54.1 |
| deadV1_FL(NtV2) (50 pM) | >9.9 | 31.0 | |||
| deadV2_FL (50 pM) | >13.4 | 19.1 | |||
| deadV2_FL(NtV1) (50 pM) | >6.5 | 18.1 | |||
| C (BRB40, 50 mM KCl) | 140 | Tyr-MTs | V1_FL (25 pM) | 1.2 | 96.7 |
| V1_FL(NtV2) (25 pM) | >10.7 | 20.8 | |||
| V2_FL (25 pM) | >12.9 | 22.9 | |||
| V2_FL(NtV1) (25 pM) | >3.9 | 42.4 |
(A) Related to Fig. 5, D and E. (B and C) Related to Fig. 6, A and B. A, B, and C represent experiments performed on different days. At least 10 microtubules were analyzed. See Materials and methods for detailed experimental conditions.
Sharing content requires targeting cookies to be enabled. Please update your cookie preferences to use this feature.