Bicelles Rich in both Sphingolipids and Cholesterol and Their Use in Studies of Membrane Proteins

James M. Hutchison; Kuo Chih Shih; Holger A. Scheidt; Sarah M. Fantin; Kristine F. Parson; George A. Pantelopulos; Haley R. Harrington; Kathleen F. Mittendorf; Shuo Qian; Richard A. Stein; Scott E. Collier; Melissa G. Chambers; John Katsaras; Markus W. Voehler; Brandon T. Ruotolo; Daniel Huster; Robert L. McFeeters; John E. Straub; Mu Ping Nieh; Charles R. Sanders

doi:10.1021/jacs.0c04669

Bicelles Rich in both Sphingolipids and Cholesterol and Their Use in Studies of Membrane Proteins

James M. Hutchison, Kuo Chih Shih, Holger A. Scheidt, Sarah M. Fantin, Kristine F. Parson, George A. Pantelopulos, Haley R. Harrington, Kathleen F. Mittendorf, Shuo Qian, Richard A. Stein, Scott E. Collier, Melissa G. Chambers, John Katsaras, Markus W. Voehler, Brandon T. Ruotolo, Daniel Huster, Robert L. McFeeters, John E. Straub, Mu Ping Nieh, Charles R. Sanders

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31 Scopus citations

Abstract

How the distinctive lipid composition of mammalian plasma membranes impacts membrane protein structure is largely unexplored, partly because of the dearth of isotropic model membrane systems that contain abundant sphingolipids and cholesterol. This gap is addressed by showing that sphingomyelin and cholesterol-rich (SCOR) lipid mixtures with phosphatidylcholine can be cosolubilized by n-dodecyl-β-melibioside to form bicelles. Small-angle X-ray and neutron scattering, as well as cryo-electron microscopy, demonstrate that these assemblies are stable over a wide range of conditions and exhibit the bilayered-disc morphology of ideal bicelles even at low lipid-to-detergent mole ratios. SCOR bicelles are shown to be compatible with a wide array of experimental techniques, as applied to the transmembrane human amyloid precursor C99 protein in this medium. These studies reveal an equilibrium between low-order oligomer structures that differ significantly from previous experimental structures of C99, providing an example of how ordered membranes alter membrane protein structure.

Original language	English
Pages (from-to)	12715-12729
Number of pages	15
Journal	Journal of the American Chemical Society
Volume	142
Issue number	29
DOIs	https://doi.org/10.1021/jacs.0c04669
State	Published - Jul 22 2020

Funding

This work was supported by National Institutes of Health (NIH) Grant Nos. RF1 AG056147 (CRS), R01 GM106672 (CRS), R01 NS095989 (CRS), and R01 GM105942 (B.T.R.). J.M.H. was supported by NIH T32 CA00958229 and by F31 AG061984. K.F.M. was supported by NIH T32 GM08320 and NSF Predoctoral Research Fellowship DGE090667. Special thanks to Dr. R. Capone for useful discussion and Dr. G. Li for proofreading. NMR instrumentation was supported by NSF (0922862), NIH (S10 RR025677), and Vanderbilt University matching funds. A portion of this research used resources at the High Flux Isotope Reactor, a Department of Energy (DOE) Office of Science User Facility operated by the Oak Ridge National Laboratory under Contract No. DE-AC05 00OR2275. The Bio-SANS instrument is supported by the Office of Biological and Environmental Research of the United States DOE. This work employed SASView 4.2.0 software, which was originally developed through the support of National Science Foundation (NSF) Award No. DMR-0520547. SasView also contains code developed with funding from the European Union Horizon 2020 program under the SINE2020 project Grant No. 654000. The LiX beamline is part of the Life Science Biomedical Technology Research resource, primarily supported by U.S. National Institute of General Medical Sciences Grant No. P41 GM111244, and by the DOE Office of Biological and Environmental Research Grant No. KP1605010, with additional support from the NIH (S10 OD012331). NSLS-II is a User Facility operated for the U.S. DOE, Office of Science, by Brookhaven National Laboratory (Contract No. DE-SC0012704). D.H. acknowledges a grant by the Deutsche Forschungsgemeinsachft (HU 720 15-2).

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Hutchison, J. M., Shih, K. C., Scheidt, H. A., Fantin, S. M., Parson, K. F., Pantelopulos, G. A., Harrington, H. R., Mittendorf, K. F., Qian, S., Stein, R. A., Collier, S. E., Chambers, M. G., Katsaras, J., Voehler, M. W., Ruotolo, B. T., Huster, D., McFeeters, R. L., Straub, J. E., Nieh, M. P., & Sanders, C. R. (2020). Bicelles Rich in both Sphingolipids and Cholesterol and Their Use in Studies of Membrane Proteins. Journal of the American Chemical Society, 142(29), 12715-12729. https://doi.org/10.1021/jacs.0c04669

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title = "Bicelles Rich in both Sphingolipids and Cholesterol and Their Use in Studies of Membrane Proteins",

abstract = "How the distinctive lipid composition of mammalian plasma membranes impacts membrane protein structure is largely unexplored, partly because of the dearth of isotropic model membrane systems that contain abundant sphingolipids and cholesterol. This gap is addressed by showing that sphingomyelin and cholesterol-rich (SCOR) lipid mixtures with phosphatidylcholine can be cosolubilized by n-dodecyl-β-melibioside to form bicelles. Small-angle X-ray and neutron scattering, as well as cryo-electron microscopy, demonstrate that these assemblies are stable over a wide range of conditions and exhibit the bilayered-disc morphology of ideal bicelles even at low lipid-to-detergent mole ratios. SCOR bicelles are shown to be compatible with a wide array of experimental techniques, as applied to the transmembrane human amyloid precursor C99 protein in this medium. These studies reveal an equilibrium between low-order oligomer structures that differ significantly from previous experimental structures of C99, providing an example of how ordered membranes alter membrane protein structure.",

author = "Hutchison, {James M.} and Shih, {Kuo Chih} and Scheidt, {Holger A.} and Fantin, {Sarah M.} and Parson, {Kristine F.} and Pantelopulos, {George A.} and Harrington, {Haley R.} and Mittendorf, {Kathleen F.} and Shuo Qian and Stein, {Richard A.} and Collier, {Scott E.} and Chambers, {Melissa G.} and John Katsaras and Voehler, {Markus W.} and Ruotolo, {Brandon T.} and Daniel Huster and McFeeters, {Robert L.} and Straub, {John E.} and Nieh, {Mu Ping} and Sanders, {Charles R.}",

note = "Publisher Copyright: {\textcopyright} 2020 American Chemical Society.",

year = "2020",

month = jul,

day = "22",

doi = "10.1021/jacs.0c04669",

language = "English",

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Hutchison, JM, Shih, KC, Scheidt, HA, Fantin, SM, Parson, KF, Pantelopulos, GA, Harrington, HR, Mittendorf, KF, Qian, S, Stein, RA, Collier, SE, Chambers, MG, Katsaras, J, Voehler, MW, Ruotolo, BT, Huster, D, McFeeters, RL, Straub, JE, Nieh, MP & Sanders, CR 2020, 'Bicelles Rich in both Sphingolipids and Cholesterol and Their Use in Studies of Membrane Proteins', Journal of the American Chemical Society, vol. 142, no. 29, pp. 12715-12729. https://doi.org/10.1021/jacs.0c04669

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AU - Hutchison, James M.

AU - Shih, Kuo Chih

AU - Scheidt, Holger A.

AU - Fantin, Sarah M.

AU - Parson, Kristine F.

AU - Pantelopulos, George A.

AU - Harrington, Haley R.

AU - Mittendorf, Kathleen F.

AU - Qian, Shuo

AU - Stein, Richard A.

AU - Collier, Scott E.

AU - Chambers, Melissa G.

AU - Katsaras, John

AU - Voehler, Markus W.

AU - Ruotolo, Brandon T.

AU - Huster, Daniel

AU - McFeeters, Robert L.

AU - Straub, John E.

AU - Nieh, Mu Ping

AU - Sanders, Charles R.

PY - 2020/7/22

Y1 - 2020/7/22

N2 - How the distinctive lipid composition of mammalian plasma membranes impacts membrane protein structure is largely unexplored, partly because of the dearth of isotropic model membrane systems that contain abundant sphingolipids and cholesterol. This gap is addressed by showing that sphingomyelin and cholesterol-rich (SCOR) lipid mixtures with phosphatidylcholine can be cosolubilized by n-dodecyl-β-melibioside to form bicelles. Small-angle X-ray and neutron scattering, as well as cryo-electron microscopy, demonstrate that these assemblies are stable over a wide range of conditions and exhibit the bilayered-disc morphology of ideal bicelles even at low lipid-to-detergent mole ratios. SCOR bicelles are shown to be compatible with a wide array of experimental techniques, as applied to the transmembrane human amyloid precursor C99 protein in this medium. These studies reveal an equilibrium between low-order oligomer structures that differ significantly from previous experimental structures of C99, providing an example of how ordered membranes alter membrane protein structure.

AB - How the distinctive lipid composition of mammalian plasma membranes impacts membrane protein structure is largely unexplored, partly because of the dearth of isotropic model membrane systems that contain abundant sphingolipids and cholesterol. This gap is addressed by showing that sphingomyelin and cholesterol-rich (SCOR) lipid mixtures with phosphatidylcholine can be cosolubilized by n-dodecyl-β-melibioside to form bicelles. Small-angle X-ray and neutron scattering, as well as cryo-electron microscopy, demonstrate that these assemblies are stable over a wide range of conditions and exhibit the bilayered-disc morphology of ideal bicelles even at low lipid-to-detergent mole ratios. SCOR bicelles are shown to be compatible with a wide array of experimental techniques, as applied to the transmembrane human amyloid precursor C99 protein in this medium. These studies reveal an equilibrium between low-order oligomer structures that differ significantly from previous experimental structures of C99, providing an example of how ordered membranes alter membrane protein structure.

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JF - Journal of the American Chemical Society

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ER -

Bicelles Rich in both Sphingolipids and Cholesterol and Their Use in Studies of Membrane Proteins

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