Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden; Science for Life Laboratory, Stockholm University, Box 1031, SE-171 21 Solna, Sweden.
J Mol Biol. 2019 Mar 15;431(6):1308-1314. doi: 10.1016/j.jmb.2019.01.043. Epub 2019 Feb 7.
We have characterized the cotranslational folding of two small protein domains of different folds-the α-helical N-terminal domain of HemK and the β-rich FLN5 filamin domain-by measuring the force that the folding protein exerts on the nascent chain when located in different parts of the ribosome exit tunnel (force-profile analysis, or FPA), allowing us to compare FPA to three other techniques currently used to study cotranslational folding: real-time FRET, photoinduced electron transfer, and NMR. We find that FPA identifies the same cotranslational folding transitions as do the other methods, and that these techniques therefore reflect the same basic process of cotranslational folding in similar ways.
我们通过测量折叠蛋白位于核糖体出口隧道不同部位时对新生链施加的力（力谱分析，或 FPA），对两种不同折叠结构的小蛋白结构域 - HemK 的α-螺旋 N 端结构域和富含β的 FLN5 细丝蛋白结构域 - 的共翻译折叠进行了表征，这使我们能够将 FPA 与目前用于研究共翻译折叠的其他三种技术进行比较：实时 FRET、光诱导电子转移和 NMR。我们发现 FPA 确定了与其他方法相同的共翻译折叠转变，因此这些技术以类似的方式反映了共翻译折叠的相同基本过程。
