Photoaffinity labeling is a technique used to study drug-target interactions in vivo by derivatizing a drug with a photocrosslinker group to create a probe molecule which, on exposure to UV light, will covalently attach to the enzyme or receptor of interest. The resulting probes can be used in affinity-based protein profiling, a proteomics technique which seeks to characterize the structure and function of related proteins and to interrogate both on- and off-target drug interactions. Recently, our lab has developed probes to study dopamine receptor D2, a G-protein coupled receptor (GPCR), by derivatizing dopamine receptor modulator drugs with structurally minimalist photocrosslinker molecules. So far, we have combined the dopamine receptor agonists ropinirole (Requip) and pramipexole (Mirapex) with a diazirine- or benzophenone-containing photoactive crosslinker to create four different probes. Each probe contains a terminal alkyne for attachment to a fluorescent azide reporter tag via a copper-catalyzed azide-alkyne cycloaddition (CuAAC) "click" reaction. The data collected from these experiments will further current understanding of binding interactions at the dopamine D2 active site, and can be used to aid in the rational design of new dopaminergic drugs. The development of these probes and subsequent microscopy experiments to test their activity in vivo will be presented.

Synthesis and in vivo testing of new clickable drug-photocrosslinker conjugates as dopamine receptor probes

ACS Fall 2021

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