title: In situ 19F NMR studies of an E. coli membrane protein.
abstract: In this report, (19)F spin incorporation in a specific site of a specific membrane protein in E. coli was accomplished via trifluoromethyl-phenylalanine ((19) F-tfmF). Site-specific (19)F chemical shifts and longitudinal relaxation times of diacylglycerol kinase (DAGK), an E. coli membrane protein, were measured in its native membrane using in situ magic angle spinning (MAS) solid state nuclear magnetic resonance (NMR). Comparing with solution NMR data of the purified DAGK in detergent micelles, the in situ MAS-NMR data illustrated that (19)F chemical shift values of residues at different membrane protein locations were influenced by interactions between membrane proteins and their surrounding lipid or lipid mimic environments, while (19)F side chain longitudinal relaxation values were probably affected by different interactions of DAGK with planar lipid bilayer versus globular detergent micelles.
authors.affiliation: Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230026, People's Republic of China.
authors.forename: Pan
authors.initials: P
authors.lastname: Shi
authors.forename: Dong
authors.initials: D
authors.lastname: Li
authors.forename: Hongwei
authors.initials: H
authors.lastname: Chen
authors.forename: Ying
authors.initials: Y
authors.lastname: Xiong
authors.forename: Yusong
authors.initials: Y
authors.lastname: Wang
authors.forename: Changlin
authors.initials: C
authors.lastname: Tian
mesh_terms: D002462:Cell Membrane / Q000737:chemistry; D003902:Detergents / Q000737:chemistry; D019852:Diacylglycerol Kinase / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry* / Q000201:enzymology; D029968:Escherichia coli Proteins / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry*; D008823:Micelles; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000737:chemistry; D025941:Protein Interaction Mapping; D012996:Solutions / Q000737:chemistry; D013997:Time Factors
chemical_list: C557397:4-(trifluoromethyl)phenylalanine; D003902:Detergents; D029968:Escherichia coli Proteins; D008051:Lipid Bilayers; D008565:Membrane Proteins; D008823:Micelles; D012996:Solutions; D010649:Phenylalanine; D019852:Diacylglycerol Kinase