Determination of the Active Site of Peptidyl-prolyl Isomerase Enzymes Using Peptide-Based Affinity Cleaving Reagents

舍帕茨博士建议使用一种新技术--蛋白质亲和力切割--来分析蛋白质折叠的机制。该方法需要合成含有蛋白质裂解试剂的双功能分子，该试剂共价连接到蛋白质结合配体上。一旦配体被结合，羟基自由基就会松动，从而在活性部位附近发生裂解。对多肽裂解部位的分析将有助于阐明活性部位和周围环境，从而解决单一氨基酸的问题。据称，它在这方面优于其他技术，如亲和标记实验。这项研究的结果有望为这一说法提供证据，并应提供将被研究的多肽-脯氨酰异构酶的有趣数据。

Dr. Schepartz proposes to use a new technique - protein affinity cleavage - to analyze the mechanism of protein folding. The method requires the synthesis of bifunctional molecules which contain a protein-cleaving reagent covalently tethered to a protein-binding ligand. Once the ligand is bound the hydroxyl radical is loosed to allow cleavage in the neighborhood of the active site. Analysis of the sites of peptide cleavage will help elucidate the active site and surround to a resolution of single amino acids. It is claimed to be superior in this regard to other techniques, such as affinity labeling experiments. The results from this study promise to provide evidence for this claim and should give interesting data on the peptidy-prolyl isomerase enzymes which will be studied.