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Bicelles Rich in both Sphingolipids and Cholesterol and Their Use in Studies of Membrane Proteins
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2020-06-23 , DOI: 10.1021/jacs.0c04669
James M Hutchison 1 , Kuo-Chih Shih 2 , Holger A Scheidt 3 , Sarah M Fantin 4 , Kristine F Parson 4 , George A Pantelopulos 5 , Haley R Harrington 6 , Kathleen F Mittendorf 7 , Shuo Qian 8 , Richard A Stein 9 , Scott E Collier 10 , Melissa G Chambers 11 , John Katsaras 12 , Markus W Voehler 13 , Brandon T Ruotolo 4 , Daniel Huster 3 , Robert L McFeeters 14 , John E Straub 5 , Mu-Ping Nieh 2 , Charles R Sanders 15
Affiliation  

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 co-solubilized 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.

中文翻译:

富含鞘脂和胆固醇的双细胞及其在膜蛋白研究中的应用

哺乳动物质膜的独特脂质组成如何影响膜蛋白结构在很大程度上尚未得到探索,部分原因是缺乏含有丰富鞘脂和胆固醇的各向同性模型膜系统。这一差距通过显示鞘磷脂和富含胆固醇 (SCOR) 脂质混合物与磷脂酰胆碱可以被正十二烷基-β-蜜二糖苷共溶以形成二聚体来解决。小角度 X 射线和中子散射以及低温电子显微镜表明,这些组件在广泛的条件下都是稳定的,即使在低脂质与洗涤剂摩尔比的情况下,也表现出理想双细胞的双层圆盘形态。SCOR bicelles 显示与广泛的实验技术兼容,如应用于该介质中的跨膜人淀粉样前体 C99 蛋白。
更新日期:2020-06-23
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