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Detection of altered protein conformations in living cells

机译:检测活细胞中蛋白质构象的改变

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The maturation, conformational stability, and the rate of in vivo degradation are specific for each protein and depend on both the intrinsic features of the protein and those of the surrounding cellular environment. While synthesis and degradation can be measured in living cells, stability and maturation of proteins are more difficult to quantify. We developed the split-ubiquitin method into a tool for detecting and analyzing changes in protein conformation. The biophysical parameter that forms the basis of these measurements is the time-averaged distance between the N terminus and C terminus of a protein. Starting from three proteins of known structure, we demonstrate the feasibility of this approach, and employ it to elucidate the effect of a previously described mutation in the protein Sec62p on its conformation in Living cells.
机译:成熟度,构象稳定性和体内降解速率对每种蛋白质都是特定的,并且取决于蛋白质的固有特征和周围细胞环境的固有特征。虽然可以在活细胞中测量合成和降解,但蛋白质的稳定性和成熟度更难量化。我们将分裂泛素方法开发为一种检测和分析蛋白质构象变化的工具。构成这些测量基础的生物物理参数是蛋白质N末端和C末端之间的时间平均距离。从已知结构的三种蛋白质开始,我们证明了这种方法的可行性,并用它来阐明蛋白质Sec62p中先前描述的突变对其在活细胞中构象的影响。

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