Ligand identification of the adenosine A2A receptor in self-assembled nanodiscs by affinity mass spectrometry

Abstract

A nanodisc is designed as a vehicle to contain GPCRs in a native-like phospholipid environment. It maintains GPCRs in a better thermostability and homogeneity state in a detergent-free buffer compared to a detergent micelle environment. Here, we established an affinity mass spectrometry-based approach to assay the interaction between the adenosine A2A receptor in the nanodisc and eight known ligands. The affinity mass spectrometry results are consistent with the previously published data for binding affinities. The combination of nanodisc and affinity MS techniques allows the identification of the GPCR ligands in a relatively stable, unliganded/apo and native state.