Development of clickable active site-directed photoaffinity probes for γ-secretase Journal Article
| Authors: | Crump, C. J.; Am Ende, C. W.; Eric Ballard, T.; Pozdnyakov, N.; Pettersson, M.; Chau, D. M.; Bales, K. R.; Li, Y. M.; Johnson, D. S. |
| Article Title: | Development of clickable active site-directed photoaffinity probes for γ-secretase |
| Abstract: | We have developed clickable active site-directed photoaffinity probes for γ-secretase which incorporate a photoreactive benzophenone group and an alkyne handle for subsequent click chemistry mediated conjugation with azide-linked reporter tags for visualization (e.g., TAMRA-azide) or enrichment (e.g., biotin-azide) of labeled proteins. Specifically, we synthesized clickable analogs of L646 (2) and L505 (3) and validated specific labeling to presenilin-1 N-terminal fragment (PS1-NTF), the active site aspartyl protease component within the γ-secretase complex. Additionally, we were able to identify signal peptide peptidase (SPP) by Western blot analysis. Furthermore, we analyzed the photo-labeled proteins in an unbiased fashion by click chemistry with TAMRA-azide followed by in-gel fluorescence detection. This approach expands the utility of γ-secretase inhibitor (GSI) photoaffinity probes in that labeled proteins can be tagged with any number of azide-linked reporters groups using a single clickable photoaffinity probe for target pull down and/or fluorescent imaging applications. © 2012 Elsevier Ltd. All rights reserved. |
| Keywords: | unclassified drug; human cell; nonhuman; validation process; animal cell; complex formation; fluorescence; drug potency; drug synthesis; gel; amino terminal sequence; western blotting; molecular probe; gamma secretase inhibitor; gamma secretase; presenilin 1; biotin; enzyme active site; photoaffinity labeling; click chemistry; aspartic proteinase; peptidase; conjugation; alzheimer's disease; alkyne; benzophenone; signal peptide peptidase; azide; γ-secretase; clickable photoprobe; l 505; l 646 |
| Journal Title: | Bioorganic & Medicinal Chemistry Letters |
| Volume: | 22 |
| Issue: | 8 |
| ISSN: | 0960-894X |
| Publisher: | Pergamon-Elsevier Science Ltd |
| Date Published: | 2012-04-15 |
| Start Page: | 2997 |
| End Page: | 3000 |
| Language: | English |
| DOI: | 10.1016/j.bmcl.2012.02.027 |
| PROVIDER: | scopus |
| PMCID: | PMC3321063 |
| PUBMED: | 22418280 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 1 May 2012" - "CODEN: BMCLE" - "Source: Scopus" |
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