title: NMR structure of Mistic, a membrane-integrating protein for membrane protein expression.
abstract: Although structure determination of soluble proteins has become routine, our understanding of membrane proteins has been limited by experimental bottlenecks in obtaining both sufficient yields of protein and ordered crystals. Mistic is an unusual Bacillus subtilis integral membrane protein that folds autonomously into the membrane, bypassing the cellular translocon machinery. Using paramagnetic probes, we determined by nuclear magnetic resonance (NMR) spectroscopy that the protein forms a helical bundle with a surprisingly polar lipid-facing surface. Additional experiments suggest that Mistic can be used for high-level production of other membrane proteins in their native conformations, including many eukaryotic proteins that have previously been intractable to bacterial expression.
authors.affiliation: Structural Biology Laboratory, Salk Institute, San Diego, CA 92037, USA.
authors.forename: Tarmo P
authors.initials: TP
authors.lastname: Roosild
authors.forename: Jason
authors.initials: J
authors.lastname: Greenwald
authors.forename: Mark
authors.initials: M
authors.lastname: Vega
authors.forename: Samantha
authors.initials: S
authors.lastname: Castronovo
authors.forename: Roland
authors.initials: R
authors.lastname: Riek
authors.forename: Senyon
authors.initials: S
authors.lastname: Choe
mesh_terms: D001412:Bacillus subtilis / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry; D018360:Crystallography, X-Ray; D004578:Electron Spin Resonance Spectroscopy; D004926:Escherichia coli; D006860:Hydrogen Bonding; D008051:Lipid Bilayers; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D008823:Micelles; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D018125:Receptors, Transforming Growth Factor beta / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D008051:Lipid Bilayers; D008565:Membrane Proteins; D008823:Micelles; D018125:Receptors, Transforming Growth Factor beta; D011994:Recombinant Proteins