title: Divergent synthesis of multifunctional molecular probes to elucidate the enzyme specificity of dipeptidic gamma-secretase inhibitors.
abstract: 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.
authors.affiliation: Laboratory of Biostructural Chemistry, Graduate School of Life Sciences, Tohoku University, Tsutsumidori-Amamiya, Aoba-ku, Sendai, Japan. hfuwa@bios.tohoku.ac.jp
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Fuwa
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Yu
authors.initials: Y
authors.lastname: Konno
authors.forename: Naoto
authors.initials: N
authors.lastname: Watanabe
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Miyashita
authors.forename: Makoto
authors.initials: M
authors.lastname: Sasaki
authors.forename: Hideaki
authors.initials: H
authors.lastname: Natsugari
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Kan
authors.forename: Tohru
authors.initials: T
authors.lastname: Fukuyama
authors.forename: Taisuke
authors.initials: T
authors.lastname: Tomita
authors.forename: Takeshi
authors.initials: T
authors.lastname: Iwatsubo
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D000818:Animals; D006367:HeLa Cells; D006801:Humans; D015335:Molecular Probes / Q000138:chemical synthesis*; D053764:Presenilin-1 / Q000378:metabolism; D011480:Protease Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D013379:Substrate Specificity / Q000187:drug effects / Q000502:physiology
chemical_list: D015335:Molecular Probes; D053764:Presenilin-1; D011480:Protease Inhibitors; D053829:Amyloid Precursor Protein Secretases