Cholesterol Increases Kinetic, Energetic, and Mechanical Stability
Cholesterol increases kinetic, energetic, and PNAS PLUS mechanical stability of the human β2-adrenergic receptor Michael Zochera, Cheng Zhangb, Søren G. F. Rasmussenb, Brian K. Kobilkab,1, and Daniel J. Müllera,1 aDepartment of Biosystems Science and Engineering, Eidgenössiche Technische Hochschule (ETH) Zurich, 4058 Basel, Switzerland; and bDepartment of Molecular and Cellular Physiology, Stanford University School of Medicine, Palo Alto, CA 94305 Edited by Wolfgang Baumeister, Max Planck Institute of Biochemistry, Martinsried, Germany, and approved October 10, 2012 (received for review July 18, 2012) The steroid cholesterol is an essential component of eukaryotic Similar to other GPCRs, cholesterol modulates the physio- membranes, and it functionally modulates membrane proteins, logical function of β2AR (26, 28). Furthermore, cholesterol and including G protein-coupled receptors. To reveal insight into how the more water-soluble cholesterol analog cholesteryl hemi- cholesterol modulates G protein-coupled receptors, we have used succinate (CHS) enhance the thermal stability of β2AR (26, 29). dynamic single-molecule force spectroscopy to quantify the me- Additionally, cholesterol facilitates interactions between GPCRs fl chanical strength and exibility, conformational variability, and and seems to be helpful in crystallizing β2AR (9). A recently kinetic and energetic stability of structural segments stabilizing published X-ray crystallography model of human β2AR showed β β the human 2-adrenergic receptor ( 2AR) in the absence and pres- cholesterol to fit into a shallow surface groove formed by ence of the cholesterol analog cholesteryl hemisuccinate (CHS). transmembrane α-helices H1, H2, H3, and H4 (30). This struc- CHS considerably increased the kinetic, energetic, and mechanical tural model unraveled possible interactions between cholesterol fi stability of almost every structural segment at suf cient magni- and β2AR.
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