| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | The gene coding for hA2AR (residues 1−316) was synthesized by Genscript and cloned into pPICZb with an N-terminal α-factor signal sequence from Sa c cha romy c e s c e r e v i s ia e (MRFPS IFTAVLFAASSLAAPVNTT - Article EDETAQIPAAVIGYSDLEDFDVAVLPSNSTNNGLLINTTIASIAA- EEGVSLERLVPRGS) , followed by hA2AR and a C-term inus b i o t i n y l a t i o n d om a i n f r om P r o p i o n i b a c t e r i um s h e rm a n i i (T S E F EN L YQGQ FGGGTG A PA PAAGGAGGKAG EG E I PA - L A G T V S K I L V E G D T V K A G Q V L V L E A M K M E E I N A PTDGKVEKVLKERDAVQGQGLIKI) for enhanced expression40 and a decaHis tag (GHHHHHHHHHGS). | Get A Quote |
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 A2A receptor (hA2AR) 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 hA2AR ligand with a terminal alkyne to serve as probe. We showed that our probe irreversibly and concentration-dependently labeled purified hA2AR. Click-ligation with a sulfonated cyanine-3 fluorophore allowed us to visualize the receptor ... More
