An Affinity-Based Probe for the Human Adenosine A_2A Receptor.

ABSTRACT: Using activity-based protein profiling (ABPP), functional proteins can be interrogated in their native environment. Despite their pharmaceutical relevance, G protein-coupled receptors (GPCRs) have been difficult to address through ABPP. In the current study, we took the prototypical human adenosine A_2A receptor (hA_2AR) as the starting point for the construction of a chemical toolbox allowing two-step affinity-based labeling of GPCRs. First, we equipped an irreversibly binding hA_2AR ligand with a terminal alkyne to serve as probe. We showed that our probe irreversibly and concentration-dependently labeled purified hA_2AR. Click-ligation with a sulfonated cyanine-3 fluorophore allowed us to visualize the receptor on SDS-PAGE. We further demonstrated

SUBMITTER: Yang X 

PROVIDER: S-EPMC6150691 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature