Identifying interactions between transmembrane helices from the adenosine A2A receptor.
The human adenosine A(2A) receptor (A(2A)R) is an integral membrane protein and a member of the G-protein-coupled receptor (GPCR) superfamily, characterized by seven transmembrane (TM) helices. Although helix-helix association in the lipid bilayer is known to be an essential step in the folding of GPCRs, the determinants of their structures, folding, and assembly in the cell membrane are poorly understood. Previous studies in our group showed that while peptides corresponding to all seven TM domains of A(2A)R form stable helical structures in detergent micelles and lipid vesicles, they display significant variability in their helical propensity. This finding suggested to us that some TM domains might need to interact with other domains to properly insert and fold in hydrophobic environments. In this study, we assessed the ability of TM peptides to interact in pairwise combinations. We analyzed peptide interactions in hydrophobic milieus using circular dichroism spectroscopy and Förster resonance energy transfer. We find that specific interactions between TM helices occur, leading to additional helical content, especially in weakly helical TM domains, suggesting that some TM domains need a partner for proper folding in the membrane. The approach developed in this study will enable complete analysis of the TM domain interactions and the modeling of a folding pathway for A(2A)R.
Thévenin
Damien
D
Department of Chemistry and Biochemistry, Delaware Biotechnology Institute, University of Delaware, Newark, Delaware 19711, USA.
Roberts
Matthew F
MF
Lazarova
Tzvetana
T
Robinson
Clifford R
CR
D002942:Circular Dichroism; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017433:Protein Structure, Secondary; D043705:Receptor, Adenosine A2A / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D010455:Peptides; D043705:Receptor, Adenosine A2A