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Photoinducible Oncometabolite Detection
Author(s) -
Kulkarni Rhushikesh A.,
Briney Chloe A.,
Crooks Daniel R.,
Bergholtz Sarah E.,
Mushti Chandrasekhar,
Lockett Stephen J.,
Lane Andrew N.,
Fan Teresa W.M.,
Swenson Rolf E.,
Marston Linehan W.,
Meier Jordan L.
Publication year - 2019
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201800651
Subject(s) - bioorthogonal chemistry , chemistry , metabolite , flow cytometry , biochemistry , computational biology , combinatorial chemistry , microbiology and biotechnology , biology , click chemistry
Dysregulated metabolism can fuel cancer by altering the production of bioenergetic building blocks and directly stimulating oncogenic gene‐expression programs. However, relatively few optical methods for the direct study of metabolites in cells exist. To address this need and facilitate new approaches to cancer treatment and diagnosis, herein we report an optimized chemical approach to detect the oncometabolite fumarate. Our strategy employs diaryl tetrazoles as cell‐permeable photoinducible precursors to nitrileimines. Uncaging these species in cells and cell extracts enables them to undergo 1,3‐dipolar cycloadditions with endogenous dipolarophile metabolites such as fumarate to form pyrazoline cycloadducts that can be readily detected by their intrinsic fluorescence. The ability to photolytically uncage diaryl tetrazoles provides greatly improved sensitivity relative to previous methods, and enables the facile detection of dysregulated fumarate metabolism through biochemical activity assays, intracellular imaging, and flow cytometry. Our studies showcase an intersection of bioorthogonal chemistry and metabolite reactivity that can be applied for biological profiling, imaging, and diagnostics.