MacKinnon Andrew L, Garrison Jennifer L, Hegde Ramanujan S, Taunton Jack
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, California 94158, USA.
J Am Chem Soc. 2007 Nov 28;129(47):14560-1. doi: 10.1021/ja076250y. Epub 2007 Nov 6.
Photoaffinity labeling is a powerful tool to identify protein targets of biologically active small molecules, yet is often limited by the size, chemical properties, and availability of photoreactive groups. We report an improved synthesis of photo-leucine, a diazirine-based photoreactive analogue of leucine, and demonstrate its incorporation into a cyclodepsipeptide inhibitor of cotranslational translocation. Photoaffinity labeling in a crude membrane fraction, followed by "click chemistry" with a rhodamine-azide reporter, enabled the identification of Sec61alpha, the structural core of the Sec61 translocation channel, as the inhibitor's target.
光亲和标记是一种用于识别生物活性小分子蛋白质靶点的强大工具，但它常常受到光反应基团的大小、化学性质和可用性的限制。我们报道了一种改进的光亮氨酸合成方法，光亮氨酸是一种基于重氮丙啶的亮氨酸光反应类似物，并证明它能掺入共翻译转运的环缩肽抑制剂中。在粗膜组分中进行光亲和标记，随后与罗丹明叠氮化物报告分子进行“点击化学”，从而能够鉴定出Sec61易位通道的结构核心Sec61α作为抑制剂的靶点。
