Conformationally selective RNA aptamers allosterically modulate the β2-adrenoceptor.

Journal Article (Journal Article)

G-protein-coupled receptor (GPCR) ligands function by stabilizing multiple, functionally distinct receptor conformations. This property underlies the ability of 'biased agonists' to activate specific subsets of a given receptor's signaling profile. However, stabilizing distinct active GPCR conformations to enable structural characterization of mechanisms underlying GPCR activation remains difficult. These challenges have accentuated the need for receptor tools that allosterically stabilize and regulate receptor function through unique, previously unappreciated mechanisms. Here, using a highly diverse RNA library combined with advanced selection strategies involving state-of-the-art next-generation sequencing and bioinformatics analyses, we identify RNA aptamers that bind a prototypical GPCR, the β2-adrenoceptor (β2AR). Using biochemical, pharmacological, and biophysical approaches, we demonstrate that these aptamers bind with nanomolar affinity at defined surfaces of the receptor, allosterically stabilizing active, inactive, and ligand-specific receptor conformations. The discovery of RNA aptamers as allosteric GPCR modulators significantly expands the diversity of ligands available to study the structural and functional regulation of GPCRs.

Full Text

Duke Authors

Cited Authors

  • Kahsai, AW; Wisler, JW; Lee, J; Ahn, S; Cahill Iii, TJ; Dennison, SM; Staus, DP; Thomsen, ARB; Anasti, KM; Pani, B; Wingler, LM; Desai, H; Bompiani, KM; Strachan, RT; Qin, X; Alam, SM; Sullenger, BA; Lefkowitz, RJ

Published Date

  • September 2016

Published In

Volume / Issue

  • 12 / 9

Start / End Page

  • 709 - 716

PubMed ID

  • 27398998

Pubmed Central ID

  • PMC4990464

Electronic International Standard Serial Number (EISSN)

  • 1552-4469

Digital Object Identifier (DOI)

  • 10.1038/nchembio.2126


  • eng

Conference Location

  • United States