Divergent synthesis of multifunctional molecular probes to elucidate the enzyme specificity of dipeptidic gamma-secretase inhibitors.
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.
Fuwa
Haruhiko
H
Laboratory of Biostructural Chemistry, Graduate School of Life Sciences, Tohoku University, Tsutsumidori-Amamiya, Aoba-ku, Sendai, Japan. hfuwa@bios.tohoku.ac.jp
Takahashi
Yasuko
Y
Konno
Yu
Y
Watanabe
Naoto
N
Miyashita
Hiroyuki
H
Sasaki
Makoto
M
Natsugari
Hideaki
H
Kan
Toshiyuki
T
Fukuyama
Tohru
T
Tomita
Taisuke
T
Iwatsubo
Takeshi
T
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
D015335:Molecular Probes; D053764:Presenilin-1; D011480:Protease Inhibitors; D053829:Amyloid Precursor Protein Secretases