Volume 451 (2013), pp. 33–44
There was an error in the concentration of the first PMA treatment given in Table 3. The corrected Table appears here.
Membrane/cytoplasmic intensity ratios were calculated from the confocal images of the multiple experiments. Values are the means±S.E.M. for at least three images for 1000 nM and 100 nM PMA-treated samples and at least two images for the 10 nM PMA-treated samples. WT, wild-type.
| Treatment . | Time (min) . | PKCθ WT . | PKCθ Y239A . | PKCθ T243A . | PKCθ W253G . | PKCθ L255G . | PKCθ Q258G . |
|---|---|---|---|---|---|---|---|
| 1000 nM PMA | 5 | 14.5±2.2 | 3.8±1.8 | 10.3±1.9 | 11.4±3.6 | 4.8±1.5 | 1.2±0.1 |
| 10 | 11.3±2.1 | 4.2±1.6 | 9.9±1.5 | 12.7±3.9 | 6.0±1.2 | 2.0±0.3 | |
| 20 | 12.1±2.2 | 5.0±0.4 | 7.4±1.5 | 13.2±5.0 | 7.2±1.9 | 2.1±0.4 | |
| 100 nM PMA | 5 | 8.1±1.2 | 1.7±0.4 | 3.6±1.1 | 1.9±0.4 | 1.3±0.1 | 1.0±0.0 |
| 10 | 6.5±0.7 | 2.3±0.8 | 3.7±0.8 | 3.0±0.7 | 1.6±0.2 | 1.1±0.0 | |
| 20 | 5.4±0.5 | 2.3±0.5 | 3.1±0.6 | 2.6±0.6 | 2.3±0.4 | 1.1±0.0 | |
| 10 nM PMA | 5 | 1.0±0.1 | 1.1±0.0 | 1.0±0.0 | 1.0±0.1 | 1.1±0.1 | 1.2±0.1 |
| 10 | 1.0±0.1 | 1.1±0.1 | 1.3±0.2 | 1.1±0.1 | 1.1±0.1 | 1.1±0.1 | |
| 20 | 1.2±0.4 | 1.0±0.0 | 1.7±0.1 | 1.2±0.2 | 1.1±0.0 | 1.1±0.0 |
| Treatment . | Time (min) . | PKCθ WT . | PKCθ Y239A . | PKCθ T243A . | PKCθ W253G . | PKCθ L255G . | PKCθ Q258G . |
|---|---|---|---|---|---|---|---|
| 1000 nM PMA | 5 | 14.5±2.2 | 3.8±1.8 | 10.3±1.9 | 11.4±3.6 | 4.8±1.5 | 1.2±0.1 |
| 10 | 11.3±2.1 | 4.2±1.6 | 9.9±1.5 | 12.7±3.9 | 6.0±1.2 | 2.0±0.3 | |
| 20 | 12.1±2.2 | 5.0±0.4 | 7.4±1.5 | 13.2±5.0 | 7.2±1.9 | 2.1±0.4 | |
| 100 nM PMA | 5 | 8.1±1.2 | 1.7±0.4 | 3.6±1.1 | 1.9±0.4 | 1.3±0.1 | 1.0±0.0 |
| 10 | 6.5±0.7 | 2.3±0.8 | 3.7±0.8 | 3.0±0.7 | 1.6±0.2 | 1.1±0.0 | |
| 20 | 5.4±0.5 | 2.3±0.5 | 3.1±0.6 | 2.6±0.6 | 2.3±0.4 | 1.1±0.0 | |
| 10 nM PMA | 5 | 1.0±0.1 | 1.1±0.0 | 1.0±0.0 | 1.0±0.1 | 1.1±0.1 | 1.2±0.1 |
| 10 | 1.0±0.1 | 1.1±0.1 | 1.3±0.2 | 1.1±0.1 | 1.1±0.1 | 1.1±0.1 | |
| 20 | 1.2±0.4 | 1.0±0.0 | 1.7±0.1 | 1.2±0.2 | 1.1±0.0 | 1.1±0.0 |
In addition, in Supplementary Table S1, the second mutant listed in the footnote was incorrect. It should read T12G, not T21G. The corrected Table appears here.
Binding affinities were calculated for PKCθC1B (PDB code 2ENZ) and PKCδC1B [1] respectively. Values are the means±S.E.M. of triplicate independent experiments. WT, wild-type.
| Protein . | Measurement . | C1B WT . | Y8A . | T12A . | W22G . | L24G . | Q27G . |
|---|---|---|---|---|---|---|---|
| PKCθ | PDBu, Kd (nM) | 0.39±0.2 | 3.32±0.34 | 0.64±0.06 | 2.95±0.19 | 4.44±0.56 | 670±150 |
| Kd (mutant)/Kd (WT) | – | 8.51 | 1.64 | 7.56 | 11.4 | 1720 | |
| PKCδ | PDBu, Kd (nM) [1,2] | 0.8±0.1 | 48±3.0* | 1.7±0.3* | 25±3.0 | Negligible | Negligible |
| Kd (mutant)/Kd (WT) | – | 60.0 | 2.1 | 31.3 | >1000 | >1000 |
| Protein . | Measurement . | C1B WT . | Y8A . | T12A . | W22G . | L24G . | Q27G . |
|---|---|---|---|---|---|---|---|
| PKCθ | PDBu, Kd (nM) | 0.39±0.2 | 3.32±0.34 | 0.64±0.06 | 2.95±0.19 | 4.44±0.56 | 670±150 |
| Kd (mutant)/Kd (WT) | – | 8.51 | 1.64 | 7.56 | 11.4 | 1720 | |
| PKCδ | PDBu, Kd (nM) [1,2] | 0.8±0.1 | 48±3.0* | 1.7±0.3* | 25±3.0 | Negligible | Negligible |
| Kd (mutant)/Kd (WT) | – | 60.0 | 2.1 | 31.3 | >1000 | >1000 |
Glycine mutants (Y8G and T12G).
