Table 3.
Genes used in biotinylation experiments
| Number | Gene | Gene family | Mr (kDa) | Biotin/unmodified |
|---|---|---|---|---|
| 1 | ARHGEF2 | G-protein-GEF | 47.2 | – |
| 2 | CBPP22 | EF hand | 25.0 | 0 |
| 3 | CDC42 | GTPase-RHO | 23.8 | N/D |
| 4 | CENTG1 | GTPase-RAS | 22.0 | – |
| 5 | DIRAS2 | GTPase-RAS | 22.1 | – |
| 6 | DIRAS1 | GTPase-RAS | 24.9 | – |
| 7 | DUSP16 | Phosphatase-Dual-Spec | 36.6 | – |
| 8 | DUSP16 | Phosphatase-Dual-Spec | 18.7 | – |
| 9 | GEM | GTPase-RAS | 23.8 | – |
| 10 | GNAI3 | GTPase-Trimeric | 42.6 | 0 |
| 11 | MAP2K2 (MEK2) | Kinase-STE | 42.3 | 0 |
| 12 | MAP2K3 (MEK3) | Kinase-STE | 37.1 | – |
| 13 | MAP2K4 (JNKK1) | Kinase-STE | 43.3 | N/D |
| 14 | MAP2K5 (MEK5) | Kinase-STE | 36.0 | N/D |
| 15 | MAP2K6 (MEK6) | Kinase-STE | 35.2 | – |
| 16 | MAP2K7 (JNKK2) | Kinase-STE | 42.9 | N/D |
| 17 | MAPK3 (ERK1) | Kinase-CMGC | 45.7 | N/D |
| 18 | MAPK6 (ERK3) | Kinase-CMGC | 40.2 | N/D |
| 19 | MAPK8 (JNK1) | Kinase-CMGC | 44.5 | – |
| 20 | MAPK9 (JNK2) | Kinase-CMGC | 46.3 | – |
| 21 | PAK4 | Kinase-STE | 66.6 | – |
| 22 | PAK7 | Kinase-STE | 38.0 | 0 |
| 23 | PAK6 | Kinase-STE | 77.4 | – |
| 24 | PPP1R12B | Phosphatase-regulatory | 39.0 | 0 |
A list of the human genes used in the experiments shown in Figs. 3 and 4. The numbering corresponds to the annotations in the figures. The molecular weights are of the proteins produced from the vector pNIC28-Bsa4, which include a His6-tag only. The C-terminal biotinylation tag adds 2.6 kDa.
The column marked Biotin/unmodified summarises the results of test expression of the C-terminal tagged proteins, comparing the yield in presence and absence of biotin in the growth medium (Fig. 3 and similar experiments). ‘0’ indicates no effect, ‘–’ indicates a reduction in yield in the presence of biotin, and N/D indicates that the yields were too low to compare.