title: Solid-state NMR spectra of lipid-anchored proteins under magic angle spinning.
abstract: Solid-state NMR is a promising tool for elucidating membrane-related biological phenomena. We achieved the measurement of high-resolution solid-state NMR spectra for a lipid-anchored protein embedded in lipid bilayers under magic angle spinning (MAS). To date, solid-state NMR measurements of lipid-anchored proteins have not been accomplished due to the difficulty in supplying sufficient amount of stable isotope labeled samples in the overexpression of lipid-anchored proteins requiring complex posttranslational modification. We designed a pseudo lipid-anchored protein in which the protein component was expressed in E. coli and attached to a chemically synthesized lipid-anchor mimic. Using two types of membranes, liposomes and bicelles, we demonstrated different types of insertion procedures for lipid-anchored protein into membranes. In the liposome sample, we were able to observe the cross-polarization and the (13)C-(13)C chemical shift correlation spectra under MAS, indicating that the liposome sample can be used to analyze molecular interactions using dipolar-based NMR experiments. In contrast, the bicelle sample showed sufficient quality of spectra through scalar-based experiments. The relaxation times and protein-membrane interaction were capable of being analyzed in the bicelle sample. These results demonstrated the applicability of two types of sample system to elucidate the roles of lipid-anchors in regulating diverse biological phenomena.
authors: Bioorganic Research Institute, Suntory Foundation for Life Sciences , 1-1-1 Wakayamadai, Shimamoto-Cho, Mishima-Gun, Osaka 618-8503, Japan.
authors: Kaoru
authors: K
authors: Nomura
authors: Erisa
authors: E
authors: Harada
authors: Kenji
authors: K
authors: Sugase
authors: Keiko
authors: K
authors: Shimamoto
mesh_terms: D002247:Carbon Isotopes / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D007553:Isotope Labeling; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D058635:Lipid-Linked Proteins / Q000737:chemistry* / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular
chemical_list: D002247:Carbon Isotopes; D008051:Lipid Bilayers; D058635:Lipid-Linked Proteins; D008081:Liposomes