A method for identifying G protein-coupled receptor dimers and their interfaces.

Jing Chen, Xin Cai, Maocai Yan, Zhengwen Wang, Zhitong Lv, Chunmei Wang

Journal: Biochimica et biophysica acta. Molecular cell research 2021;1868(1):118887

PMID: 33075383

Free Full Text on Elsevier Science View on PubMed

Abstract

The G protein-coupled receptor (GPCR) dimer interface plays an important role in the formation and stabilization of the dimer. Therefore, identifying the potential receptor-receptor interface is an important part of studying GPCRs. Various strategies have been employed to study the GPCR dimer interface and explore its functional significance, but experimental methods lack robustness and calculations are laborious. Herein, we report a combined optimized experimental and calculation approach for identifying and structurally characterizing GPCR dimer interfaces, and constructing atomic resolution models. Using a transmembrane domain (TM) peptide containing a human immunodeficiency virus trans-acting transcriptional activator (HIV-TAT) protein transduction motif, matrix-assisted laser desorption tandem time-of-flight mass spectrometry (MALDITOF-MS), and bioluminescence resonance energy transfer (BRET), we successfully identified Apelin receptor (APJ)/Nociceptin receptor 1 (ORL1) and APJ/Vasopressin receptor 2 (V2R) heterodimer interfaces, and two key sites mediating dimerization. This method can identify dimer interfaces of GPCR homodimers and heterodimers.

Address: Neurobiology Key Laboratory of Jining Medical University, Jining 272067, China; Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry CV4 7AL, UK. Electronic address: [email protected].; School of Basic Medicine, Weifang Medical University, Weifang 261053, China.; School of Pharmacy, Jining Medical University, Rizhao 276826, China.; Neurobiology Key Laboratory of Jining Medical University, Jining 272067, China.

DOI: 10.1016/j.bbamcr.2020.118887

Link outs

Free resources

Full Text Sources:

https://linkinghub.elsevier.com/retrieve/pii/S0167...

Other Literature Sources:

https://scite.ai/reports/33075383

MeSH Terms: Amino Acid Motifs, Cell Membrane, Dimerization, GTP-Binding Proteins, HEK293 Cells, Humans, Models, Molecular, Protein Conformation, Protein Domains, Protein Multimerization, Receptors, G-Protein-Coupled, Recombinant Fusion Proteins, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Survivin, tat Gene Products, Human Immunodeficiency Virus