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...
FKBP-DTag技术的基本原理是利用FKBP和FRB之间的结合能力,将FKBP和FRB标记在需要研究的蛋白质上,并通过添加小分子化合物Rapamycin来诱导FKBP和FRB的结合。Rapamycin是一种免疫抑制剂,能够与FKBP和FRB形成三聚体复合物。当FKBP和FRB标记的蛋白质分别与Rapamycin结合时,它们会相互吸引并形成复合物,从而实现蛋白质的相互作用。
Figure 7.2.mTOR domain structure, complexes and downstream signaling. mTOR is a 289-kDa serine/threonine protein kinase, containing multiple domains which include the FAT domain, the FKBP12-rapamycin binding domain (FRB), the kinase domain, the FATC domain and at least 20 HEAT repeats, which pr...
These studies suggest that rapamycin's ability to bind to FRB, and by extension to mTOR, in the absence of FKBP is of little consequence under physiological conditions. Furthermore, protein-protein interactions at the FKBP12-FRB interface play a role in the stability of the ternary complex....
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 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...
Rapamycin was docked in based on the FKBP52:rapamycin crystal structure (PDB ID 4DRJ) and FK506 was docked in based on the FKBP52:FK506:FRB crystal structure (PDB ID 4LAX). Extended Data Fig. 5 Analysis of the GR:Hsp90:FKBP52 Structure. a, Expression of human FKBP52 or FKBP52...
Rapamycin-induced oligomer formation system of FRB-FKBP fusion proteins Most proteins form larger protein complexes and perform multiple functions in the cell. Thus, artificial regulation of protein complex formation controls t... T Inobe,N Nukina - 《Journal of Bioscience & Bioengineering》 被引量...
Rapamycin was docked in based on the FKBP52:rapamycin crystal structure (PDB ID 4DRJ) and FK506 was docked in based on the FKBP52:FK506:FRB crystal structure (PDB ID 4LAX). Extended Data Fig. 5 Analysis of the GR:Hsp90:FKBP52 Structure. a, Expression of human FKBP52 or FKBP52...
Here we show that none of the FKBPs from the model plant Arabidopsis (AtFKBPs) is able to form a ternary complex with the FRB domain of AtTOR in the presence of rapamycin in a two hybrid system. An antibody has been raised against the AtTOR protein and binding of recombinant yeast Sc...