title: Development of Sulfonamide Photoaffinity Inhibitors for Probing Cellular γ-Secretase.
abstract: γ-Secretase is a multiprotein complex that catalyzes intramembrane proteolysis associated with Alzheimer's disease and cancer. Here, we have developed potent sulfonamide clickable photoaffinity probes that target γ-secretase in vitro and in cells by incorporating various photoreactive groups and walking the clickable alkyne handle to different positions around the molecule. We found that benzophenone is preferred over diazirine as a photoreactive group within the sulfonamide scaffold for labeling γ-secretase. Intriguingly, the placement of the alkyne at different positions has little effect on probe potency but has a significant impact on the efficiency of labeling of γ-secretase. Moreover, the optimized clickable photoprobe, 163-BP3, was utilized as a cellular probe to effectively assess the target engagement of inhibitors with γ-secretase in primary neuronal cells. In addition, biotinylated 163-BP3 probes were developed and used to capture the native γ-secretase complex in the 3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propanesulfonate (CHAPSO) solubilized state. Taken together, these next generation clickable and biotinylated sulfonamide probes offer new tools to study γ-secretase in biochemical and cellular systems. Finally, the data provide insights into structural features of the sulfonamide inhibitor binding site in relation to the active site and into the design of clickable photoaffinity probes.
authors.affiliation: Chemical Biology Program, Memorial Sloan Kettering Cancer Center , 1275 York Avenue, New York, New York 10065, United States.
authors.forename: Christina J
authors.initials: CJ
authors.lastname: Crump
authors.affiliation: Pfizer Worldwide Research and Development , Worldwide Medicinal Chemistry, Cambridge, Massachusetts 02139, United States.
authors.forename: Heather E
authors.initials: HE
authors.lastname: Murrey
authors.affiliation: Pfizer Worldwide Research and Development , Worldwide Medicinal Chemistry Groton, Connecticut 06340, United States.
authors.forename: T Eric
authors.initials: TE
authors.lastname: Ballard
authors.affiliation: Pfizer Worldwide Research and Development , Worldwide Medicinal Chemistry Groton, Connecticut 06340, United States.
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Am Ende
authors.affiliation: Chemical Biology Program, Memorial Sloan Kettering Cancer Center , 1275 York Avenue, New York, New York 10065, United States.
authors.forename: Xianzhong
authors.initials: X
authors.lastname: Wu
authors.affiliation: Chemical Biology Program, Memorial Sloan Kettering Cancer Center , 1275 York Avenue, New York, New York 10065, United States.
authors.forename: Natalya
authors.initials: N
authors.lastname: Gertsik
authors.affiliation: Pfizer Worldwide Research and Development , Worldwide Medicinal Chemistry, Cambridge, Massachusetts 02139, United States.
authors.forename: Douglas S
authors.initials: DS
authors.lastname: Johnson
authors.affiliation: Chemical Biology Program, Memorial Sloan Kettering Cancer Center , 1275 York Avenue, New York, New York 10065, United States.
authors.forename: Yue-Ming
authors.initials: YM
authors.lastname: Li
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism*; D016229:Amyloid beta-Peptides / Q000378:metabolism; D000818:Animals; D001577:Benzophenones / Q000378:metabolism; D001665:Binding Sites / Q000187:drug effects; D019921:Biotinylation; D020134:Catalytic Domain / Q000187:drug effects*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D002793:Cholic Acids / Q000378:metabolism; D003978:Diazomethane / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology*; D010446:Peptide Fragments / Q000378:metabolism; D019920:Photoaffinity Labels; D053764:Presenilin-1 / Q000378:metabolism; D013379:Substrate Specificity; D013449:Sulfonamides / Q000494:pharmacology*
chemical_list: D016229:Amyloid beta-Peptides; D001577:Benzophenones; D002793:Cholic Acids; D010446:Peptide Fragments; D019920:Photoaffinity Labels; D053764:Presenilin-1; D013449:Sulfonamides; C075222:amyloid beta-protein (1-42); D003978:Diazomethane; C047723:benzophenone; C048531:chapso; D053829:Amyloid Precursor Protein Secretases
keywords: Alzheimer’s disease; benzophenone; click walking; clickable photoaffinity probe; diazirine; presenilin; β-amyloid