期刊
JOURNAL OF BIOMOLECULAR NMR
卷 61, 期 3-4, 页码 275-286出版社
SPRINGER
DOI: 10.1007/s10858-014-9893-4
关键词
Nanodisc; NMR; Sedimentation; Membrane protein; Ail; Yersinia
资金
- National Institutes of Health [GM100265, P41 EB002031, P30 CA030199]
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.
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