Table 1 List of protein targets whose PTM level was detected with ImmunoBRET, and the corresponding experimental values obtained in this study.
From: Monitoring phosphorylation and acetylation of CRISPR-mediated HiBiT-tagged endogenous proteins
Target | Size, kDa | PTM | Ligand | EC50 | 95% CI | Inhibitor | IC50, nM | 95% CI |
|---|---|---|---|---|---|---|---|---|
EGFR-HiBiT | 170 | p-EGFR (Y1045) | EGF | 7.819* ng/mL | 5.803–10.69* | Gefitinib | 30.19 | 26.62–33.74 |
p-EGFR (Y1068) | 11.73* ng/mL | 9.576–14.80* | 21.57 | 15.21–31.12 | ||||
p-EGFR (Y1173) | 23.32* ng/mL | 17.27–36.96* | 12.07 | 7.975–18.47 | ||||
P-Tyr-1000 MultiMab | 14.61* ng/mL | 12.90–16.74* | 21.39 | 17.22–26.78 | ||||
MET-HiBiT | 84 | p-MET (Y1349) | HGF | 48.12 ng/mL | 34.75–73.93 | Crizotinib | 25.4 | 21.51–32.41 |
STAT3-HiBiT(K562) | 83–88 | p-STAT3 (Y705) | IL-27 | 11.38 ng/mL | 9.437–13.98 | Ruxolitinib | 6.919 | 4.928–9.338 |
STAT3-HiBiT(LNCaP) | IL-6 | 12.81 ng/mL | 11.83–28.43 | 19.29 | 10.26–46.04 | |||
MAPK8-HiBiT | 44–48 | p-SAPK/JNK (T183/Y185) | TNF-α | 1.617 ng/mL | 0.6786–2.858 | SP 600125 | 22,910 | 17,000–30,920 |
Anisomycin | 53.11 nM | 4.884–65.50 | N/A | N/A | ||||
HiBiT-H3F3A | 15 | Ac-H3K9 | Trichostatin A | 0.4583 µM | 0.3436–0.6040 | N/A | N/A | N/A |
Ac-H3K27 | 0.5363 µM | 0.4052–0.7412 | N/A | N/A | N/A |
