Table 2.
Diffusion Coefficients and Mobility of CLIMP-63 Constructs
| Constructs | D | Mobility | n |
|---|---|---|---|
| 10−10 cm2s−1 | |||
| wt-GFP | 1.5 ± 0.4 | + | 5 |
| Δlumen-GFP | 8.6 ± 3.3 | +++ | 6 |
| Δ24–101-GFP | 3.1 ± 2.1 | + | 12 |
| Δ24–101Δlumen-GFP | 13.7 ± 2.2 | +++ | 4 |
| GFP-Δcytoplasmic | 8.9 ± 2.4 | ++ | 9 |
| GFP-Δcytoplasmic double luminal | 2.7 ± 1.2 | − | 4 |
| GFP-Δcytoplasmic-Δ511–601 | 7.8 ± 4.2 | ++ | 5 |
| GFP-Δcytoplasmic-Δ428–601 | 16.4 ± 8.2 | +++ | 4 |
| GFP-Δcytoplasmic-Δ354–601 | 17.8 ± 2.7 | +++ | 5 |
| GFP-Δcytoplasmic-Δ218–601 | 17.8 ± 4.9 | +++ | 5 |
| GFP-Δcytoplasmic-Δ246–492 | 6.7 ± 2.5 | ++ | 8 |
| GFP-Δcytoplasmic-Δ132–354 | 10.8 ± 5.5 | +++ | 6 |
| P450 2C2-GFP | 13.1 ± 6.6 | +++ | 6 |
| Sec61β-GFP | 7.8 ± 1.2 | +++ | 6 |
| VSV-G-GFP | 5.9 ± 1.3 | +++ | 5 |
FRAP analysis of GFP fusion constructs reveal strong influence of the luminal segment on diffusion. Deletion of the luminal segment increases diffusion and mobility (Δlumen-GFP) in contrast to deletion of the microtubule binding domain (Δ24–101-GFP), which increases diffusion but not mobility. Note that deletions of the COOH-terminal part have a higher diffusion coefficient. D, diffusion coefficient; n, number of independent experiments. Mobility is measured as a percentage of initial fluorescence: 0–25%, −; 25–50%,+; 50–75%, ++; 75–100%, +++.