Muralidharan Vasant, Muir Tom W
Laboratory of Synthetic Protein Chemistry, The Rockefeller University, New York, New York 10021, USA.
Nat Methods. 2006 Jun;3(6):429-38. doi: 10.1038/nmeth886.
Biophysical techniques such as fluorescence spectroscopy and nuclear magnetic resonance (NMR) spectroscopy provide a window into the inner workings of proteins. These approaches make use of probes that can either be naturally present within the protein or introduced through a labeling procedure. In general, the more control one has over the type, location and number of probes in a protein, then the more information one can extract from a given biophysical analysis. Recently, two related approaches have emerged that allow proteins to be labeled with a broad range of physical probes. Expressed protein ligation (EPL) and protein trans-splicing (PTS) are both intein-based approaches that permit the assembly of a protein from smaller synthetic and/or recombinant pieces. Here we provide some guidelines for the use of EPL and PTS, and highlight how the dovetailing of these new protein chemistry methods with standard biophysical techniques has improved our ability to interrogate protein function, structure and folding.
诸如荧光光谱法和核磁共振（NMR）光谱法等生物物理技术为了解蛋白质的内部运作提供了一扇窗口。这些方法利用的探针既可以天然存在于蛋白质中，也可以通过标记程序引入。一般来说，对蛋白质中探针的类型、位置和数量控制得越多，那么从给定的生物物理分析中能够提取的信息就越多。最近，出现了两种相关方法，可使蛋白质用多种物理探针进行标记。表达蛋白连接（EPL）和蛋白质反式剪接（PTS）都是基于内含肽的方法，它们允许从较小的合成和/或重组片段组装蛋白质。在此，我们提供一些使用EPL和PTS的指导原则，并强调这些新的蛋白质化学方法与标准生物物理技术的结合如何提高了我们探究蛋白质功能、结构和折叠的能力。
