Cholesterol increases kinetic, energetic, and mechanical stability of the human β2-adrenergic receptor

TY - JOUR

T1 - Cholesterol increases kinetic, energetic, and mechanical stability of the human β2-adrenergic receptor

AU - Zocher, Michael

AU - Zhang, Cheng

AU - Rasmussen, Søren Gøgsig Faarup

AU - Kobilka, Brian K

AU - Müller, Daniel J

PY - 2012/12/11

Y1 - 2012/12/11

N2 - 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.

AB - 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.

KW - Animals

KW - Biomechanical Phenomena

KW - Cholesterol Esters

KW - Energy Metabolism

KW - Humans

KW - Kinetics

KW - Microscopy, Atomic Force

KW - Models, Molecular

KW - Protein Conformation

KW - Protein Interaction Mapping

KW - Protein Stability

KW - Proteolipids

KW - Receptors, Adrenergic, beta-2

KW - Recombinant Proteins

KW - Sf9 Cells

KW - Spodoptera

KW - Unfolded Protein Response

U2 - 10.1073/pnas.1210373109

DO - 10.1073/pnas.1210373109

M3 - Journal article

C2 - 23151510

SN - 0027-8424

VL - 109

SP - E3463–E3472

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 50

ER -