Extended Data Table 2 |. Testing of unique cysteine for labeling DynCLM at its MTBD.
Protein expression and motility properties of DynCLM constructs mutated with a single surface-exposed cysteine at the MTBD, compared to WT Dyn314kDa. Assays were performed in 1 mM ATP. The most processive and well-expressing motor (Q3231C) was used for the tracking assays. The denaturing gel used for quantifying protein expression levels is shown in Extended Data Fig. 1.
| Construct Number | Construct Name | Expression Level | Average velocity (nm/s) | % processive motors landed on MTs |
|---|---|---|---|---|
| - | Wildtype | High | 80 | - |
| 1 | Q3147C | Low | 10 | >90 |
| 2 | H3169C | High | 80 | 50 |
| 3 | Q3179C | High | 70 | 50 |
| 4 | K3234C | - | - | - |
| 5 | K3242C | Medium | 50 | 50 |
| 6 | S3097C | Low | 100 | 25 |
| 7 | S3190C | Medium | 78.6 | 25 |
| 8 | K3182C | Medium | 73.3 | 25 |
| 9 | Q3211C | Medium | 76.6 | 25 |
| 10 | Q3231C | Medium | 80 | >90 |
| 11 | H3175C | Low | 83.3 | 50 |
| 12 | E3107C | Low | 58 | 25 |