Mapping the binding site of BMS-708163 on γ-secretase with cleavable photoprobes Journal Article
| Authors: | Gertsik, N.; am Ende, C. W.; Geoghegan, K. F.; Nguyen, C.; Mukherjee, P.; Mente, S.; Seneviratne, U.; Johnson, D. S.; Li, Y. M. |
| Article Title: | Mapping the binding site of BMS-708163 on γ-secretase with cleavable photoprobes |
| Abstract: | γ-Secretase, a four-subunit transmembrane aspartic proteinase, is a highly valued drug target in Alzheimer's disease and cancer. Despite significant progress in structural studies, the respective molecular mechanisms and binding modes of γ-secretase inhibitors (GSIs) and modulators (GSMs) remain uncertain. Here, we developed biotinylated cleavable-linker photoprobes based on the BMS-708163 GSI to study its interaction with γ-secretase. Comparison of four cleavable linkers indicated that the hydrazine-labile N-1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl (Dde) linker was cleaved most efficiently to release photolabeled and affinity-captured presenilin-1 (PS1), the catalytic subunit of γ-secretase. Peptide mapping showed that the BMS-708163-based probe photoinserted at L282 of PS1. This insertion site was consistent with the results of molecular dynamics simulations of the γ-secretase complex with inhibitor. Taken together, this work reveals the binding site of a GSI and offers insights into the mechanism of action of this class of inhibitors. © 2017 Elsevier Ltd |
| Keywords: | photoaffinity labeling; peptide mapping; alzheimer's disease; molecular dynamics simulations; cleavable linkers |
| Journal Title: | Cell Chemical Biology |
| Volume: | 24 |
| Issue: | 1 |
| ISSN: | 2451-9456 |
| Publisher: | Cell Press |
| Date Published: | 2017-01-19 |
| Start Page: | 3 |
| End Page: | 8 |
| Language: | English |
| DOI: | 10.1016/j.chembiol.2016.12.006 |
| PROVIDER: | scopus |
| PUBMED: | 28065657 |
| PMCID: | PMC5516958 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 2 February 2017 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work