之后，研究人员引入Rapa诱导的FKBP·FRB系统，分别构建了NUb-FRBE3(63)和FKBP-CUbPCNA融合蛋白，体外实验同样证实，Rapa诱导能够促进底物的K63链接多聚泛素化修饰。同时，为了降低由Nub和CUb之间亲和性引起的背景泛素化活性，研究人员在NUb中引入了一个点突变I13A，这个突变质粒标记为NUa，并将融合质粒NUa-HA-FRB称为...
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想要用BRET方法来鉴定FKBP12与FRB的相互作用。但是看得文献上都是需要共转至细胞里进行实验。有没有人...
The present study exemplifies the PPI in vitro inside a unique single-chain probe, named TP2.4, which carries a full-length artificial luciferase (ALuc) sandwiched in between two model proteins of interest, e.g., FKBP and FRB, expressed in E. coli, and purified. We found that the TP2.4...
The structure of the FKBP12–rapamycin–FRB ternary complex has now been refined at 2.2 Å resolution. The cell-cycle arrest agent rapamycin binds FK506-binding protein (FKBP12) and the FKBP12–rapamycin binding (FRB) domain of FKBP12–rapamycin associated protein (FRAP) simultaneously, and...
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research papers Re 庐 ned structure of the FKBP12 卤 rapamycin 卤 FRB 脢 resolution ternary complex at 2 . 2 A research papersLiang, JunChoi, JungwonClardy, Jon
Here, we engineered a rapamycin-induced dimerization system of Fas consisting of FKBP and FRB proteins. Treatment of rapamycin specifically induces cellular apoptosis. In neurons and cells with high c-FLIP expression, rapamycin-induced Fas activation triggered the activation of the non-apoptotic pathway...
The FKBP12·rapamycin complex, however, binds to FRB 2000-fold more tightly (Kd = 12 ± 0.8 nM) than rapamycin alone. No interaction between FKBP and FRB was detected in the absence of rapamycin. These studies suggest that rapamycin's ability to bind to FRB, and by extension to mTOR, ...