Fuwa Haruhiko, Takahashi Yasuko, Konno Yu, Watanabe Naoto, Miyashita Hiroyuki, Sasaki Makoto, Natsugari Hideaki, Kan Toshiyuki, Fukuyama Tohru, Tomita Taisuke, Iwatsubo Takeshi
Laboratory of Biostructural Chemistry, Graduate School of Life Sciences, Tohoku University, Tsutsumidori-Amamiya, Aoba-ku, Sendai, Japan.
ACS Chem Biol. 2007 Jun 15;2(6):408-18. doi: 10.1021/cb700073y. Epub 2007 May 25.
Divergent synthesis of multifunctional molecular probes based on caprolactam-derived dipeptidic gamma-secretase inhibitors (GSIs), Compound E (CE) and LY411575 analogue (DBZ), was efficiently accomplished by means of Cu(I)-catalyzed azide/alkyne fusion reaction. Photoaffinity labeling experiments using these derivatives coupled to photoactivatable and biotin moieties provided direct evidence that the molecular targets of CE and DBZ are the N-terminal fragment of presenilin 1 within the gamma-secretase complex. Moreover, these photoprobes directly targeted signal peptide peptidase. These data suggest that the divergent synthesis of molecular probes has been successfully applied to characterize the interaction of GSIs with their molecular targets and define the structural requirements for inhibitor binding to intramembrane-cleaving proteases.
基于己内酰胺衍生的二肽γ-分泌酶抑制剂（GSIs）、化合物E（CE）和LY411575类似物（DBZ）的多功能分子探针的发散合成，通过铜（I）催化的叠氮化物/炔烃融合反应高效完成。使用与光活化和生物素部分偶联的这些衍生物进行的光亲和标记实验提供了直接证据，表明CE和DBZ的分子靶点是γ-分泌酶复合物中早老素1的N端片段。此外，这些光探针直接靶向信号肽肽酶。这些数据表明，分子探针的发散合成已成功应用于表征GSIs与其分子靶点的相互作用，并确定抑制剂与膜内裂解蛋白酶结合的结构要求。
