Abstract
The steroid cholesterol is an essential component of eukaryotic membranes, and it functionally modulates membrane proteins, including G protein-coupled receptors. To reveal insight into how cholesterol modulates G protein-coupled receptors, we have used dynamic single-molecule force spectroscopy to quantify the mechanical strength and flexibility, conformational variability, and kinetic and energetic stability of structural segments stabilizing the human β(2)-adrenergic receptor (β(2)AR) in the absence and presence of the cholesterol analog cholesteryl hemisuccinate (CHS). CHS considerably increased the kinetic, energetic, and mechanical stability of almost every structural segment at sufficient magnitude to alter the structure and functional relationship of β(2)AR. One exception was the structural core segment of β(2)AR, which establishes multiple ligand binding sites, and its properties were not significantly influenced by CHS.
| Original language | English |
|---|---|
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 109 |
| Issue number | 50 |
| Pages (from-to) | E3463–E3472 |
| ISSN | 0027-8424 |
| DOIs | |
| Publication status | Published - 11 Dec 2012 |
| Externally published | Yes |
Keywords
- Animals
- Biomechanical Phenomena
- Cholesterol Esters
- Energy Metabolism
- Humans
- Kinetics
- Microscopy, Atomic Force
- Models, Molecular
- Protein Conformation
- Protein Interaction Mapping
- Protein Stability
- Proteolipids
- Receptors, Adrenergic, beta-2
- Recombinant Proteins
- Sf9 Cells
- Spodoptera
- Unfolded Protein Response