title: Solid-state NMR of the Yersinia pestis outer membrane protein Ail in lipid bilayer nanodiscs sedimented by ultracentrifugation.
abstract: Solid-state NMR studies of sedimented soluble proteins has been developed recently as an attractive approach for overcoming the size limitations of solution NMR spectroscopy while bypassing the need for sample crystallization or precipitation (Bertini et al. Proc Natl Acad Sci USA 108(26):10396-10399, 2011). Inspired by the potential benefits of this method, we have investigated the ability to sediment lipid bilayer nanodiscs reconstituted with a membrane protein. In this study, we show that nanodiscs containing the outer membrane protein Ail from Yersinia pestis can be sedimented for solid-state NMR structural studies, without the need for precipitation or lyophilization. Optimized preparations of Ail in phospholipid nanodiscs support both the structure and the fibronectin binding activity of the protein. The same sample can be used for solution NMR, solid-state NMR and activity assays, facilitating structure-activity correlation experiments across a wide range of timescales.
authors.affiliation: Sanford-Burnham Medical Research Institute, 10901 North Torrey Pines Road, La Jolla, CA, 92037, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Ding
authors.forename: L Miya
authors.initials: LM
authors.lastname: Fujimoto
authors.forename: Yong
authors.initials: Y
authors.lastname: Yao
authors.forename: Francesca M
authors.initials: FM
authors.lastname: Marassi
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000648:ultrastructure*; D005353:Fibronectins / Q000378:metabolism; D008051:Lipid Bilayers / Q000737:chemistry*; D008823:Micelles; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010743:Phospholipids / Q000737:chemistry; D010767:Phosphorylcholine / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D014461:Ultracentrifugation / Q000379:methods*; D037521:Virulence Factors / Q000737:chemistry; D015010:Yersinia pestis / Q000378:metabolism
chemical_list: C526543:Ail protein, Yersinia pestis; D001425:Bacterial Outer Membrane Proteins; D005353:Fibronectins; D008051:Lipid Bilayers; D008823:Micelles; D010743:Phospholipids; D037521:Virulence Factors; D010767:Phosphorylcholine