title: Cellular solid-state nuclear magnetic resonance spectroscopy.
abstract: Decrypting the structure, function, and molecular interactions of complex molecular machines in their cellular context and at atomic resolution is of prime importance for understanding fundamental physiological processes. Nuclear magnetic resonance is a well-established imaging method that can visualize cellular entities at the micrometer scale and can be used to obtain 3D atomic structures under in vitro conditions. Here, we introduce a solid-state NMR approach that provides atomic level insights into cell-associated molecular components. By combining dedicated protein production and labeling schemes with tailored solid-state NMR pulse methods, we obtained structural information of a recombinant integral membrane protein and the major endogenous molecular components in a bacterial environment. Our approach permits studying entire cellular compartments as well as cell-associated proteins at the same time and at atomic resolution.
authors.affiliation: Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Marie
authors.initials: M
authors.lastname: Renault
authors.forename: Ria
authors.initials: R
authors.lastname: Tommassen-van Boxtel
authors.forename: Martine P
authors.initials: MP
authors.lastname: Bos
authors.forename: Jan Andries
authors.initials: JA
authors.lastname: Post
authors.forename: Jan
authors.initials: J
authors.lastname: Tommassen
authors.forename: Marc
authors.initials: M
authors.lastname: Baldus
mesh_terms: D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism; D008070:Lipopolysaccharides / Q000737:chemistry / Q000378:metabolism; D008074:Lipoproteins / Q000737:chemistry / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008969:Molecular Sequence Data; D010457:Peptidoglycan / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D011510:Proteolipids / Q000737:chemistry / Q000378:metabolism
chemical_list: D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D008070:Lipopolysaccharides; D008074:Lipoproteins; C099932:Lpp protein, E coli; D010457:Peptidoglycan; D011510:Proteolipids; C020485:proteoliposomes