Selective substitution of tryptophan analogs in recombinant proteins for fluorescence studies of protein-protein interactions

几种生物和物种的基因组现在已经被揭示。这些基因组的基因产物的初级序列已经被鉴定，其中大部分是蛋白质，许多结构和功能未知。蛋白质之间的相互作用和复合体，或与DNA或RNA的相互作用和复合体如何导致生理功能或事件，是人们非常感兴趣的。研究表明，蛋白质荧光性质的信息量不仅可以揭示蛋白质复合体相互作用链段的重要结构和动力学信息，而且可以通过非相互作用链段的构象结构揭示蛋白质复合体的重要结构和动力学信息。这些荧光研究通常利用芳香氨基酸色氨酸(Trp)的固有荧光。然而，当蛋白质复合体中有多个Trp残基时，不可能揭示单个Trp残基的荧光性质和属性变化。这项研究计划描述了单个色氨酸残基如何被光谱上不同的色氨酸类似物，如5-羟基Trp，利用体外方法和无细胞蛋白质合成来取代。

The genomes of several organisms and species have now been revealed. The primary sequences of the gene products of these genomes, the majority being proteins, many of unknown structure and function, have been identified. There is considerable interest in how the interactions and complexes of proteins with one another or with DNA or RNA result in physiological functionality or events. It has been shown that the information content of the fluorescence properties of proteins can reveal important structural and dynamic information of not only the interacting segments of protein complexes by also the conformational structures of the non-interacting segments. These fluorescence studies often take advantage of the intrinsic fluorescence of the aromatic amino acid, tryptophan (Trp). However, when there are a number of Trp residues in the protein complex it is not possible to unravel the fluorescence properties and attribute changes to a single Trp residue. This research proposal describes how a single Trp residue may be substituted by a spectroscopically distinct tryptophan analogue such as 5-hydroxyTrp using in vitro methods and cell free protein synthesis.