Single Stabilizing Point Mutation Enables High‐Resolution Co‐Crystal Structures of the Adenosine A2A Receptor with Preladenant Conjugates. Issue 22 (24th March 2022)

• Record Type: Journal Article
• Authors: Claff, Tobias; Klapschinski, Tim A.; Tiruttani Subhramanyam, Udaya K.; Vaaßen, Victoria J.; Schlegel, Jonathan G.; Vielmuth, Christin; Voß, Jan H.; Labahn, Jörg; Müller, Christa E.
• Is Part Of: Angewandte Chemie international edition. Volume 61:Issue 22(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-03-24
• Subjects: Adenosine A2A Receptor -- Cancer -- G Protein-Coupled Receptor (GPCR) -- Preladenant Conjugates -- Protein Structures; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.202115545
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: The G protein‐coupled adenosine A2A receptor (A2A AR) is an important new (potential) drug target in immuno‐oncology, and for neurodegenerative diseases. Preladenant and its derivatives belong to the most potent A2A AR antagonists displaying exceptional selectivity. While crystal structures of the human A2A AR have been solved, mostly using the A2A ‐StaR2 protein that bears 9 point mutations, co‐crystallization with Preladenant derivatives has so far been elusive. We developed a new A2A AR construct harboring a single point mutation (S91 3.39 K) which renders it extremely thermostable. This allowed the co‐crystallization of two novel Preladenant derivatives, the polyethylene glycol‐conjugated (PEGylated) PSB‐2113, and the fluorophore‐labeled PSB‐2115. The obtained crystal structures (2.25 Å and 2.6 Å resolution) provide explanations for the high potency and selectivity of Preladenant derivatives. They represent the first crystal structures of a GPCR in complex with PEG‐ and fluorophore‐conjugated ligands. The applied strategy is predicted to be applicable to further class A GPCRs. Abstract : A super‐stable adenosine A2A receptor (A2A AR) containing a single point mutation in the transmembrane regions was developed by imitating an allosteric Na + through introduction of a basic amino acid residue. This allowed the determination of high‐resolution co‐crystal structures with newly designed PEGylated and fluorophore‐labeled Preladenant derivatives, which belong to the most potent A2A AR antagonists endowed with outstanding selectivity.