The study of 5-Hydroxytryptamine 2B receptor-nanodisc self-assembling and its ligand binding activity

Objective To explore the application of nanodisc in functional and drug discovery research of G protein-coupled receptor (GPCR). Methods The purified recombinant 5-Hydroxytryptamine 2B receptor (5-HT2BR) was reconstituted into nanodisc complex. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and size exclution chromatography were performed to evaluate the reconstitution reaction, followed by the use of surface plasmon resonance to validate the ligand-binding activity of 5-HT2BR after reconstitution. Results 5-HT2BR was effectively self-assembled into nanodisc while maintained its binding activity toward the antagonist SB204741. Conclusions The presented study provided potential application of 5-HT2BR-nanodisc for the development of subtype-selective drugs against 5-HT2BR and the fundamental of utilizing nanodisc for GPCR structural and functional studies as well as drug discovery.