Abstract Oriented solid-state NMR is the most direct methodology to obtain the orientation of membrane proteins with respect to the lipid bilayer. The method consists of measuring 1H-15N dipolar couplings (DC) and 15N anisotropic chemical shifts (CSA) for membrane proteins that are uniformly aligned with respect to the membrane bilayer. A significant advantage of this approach is that tilt and azimuthal (rotational) angles of the protein domains can be directly derived from analytical expression of DC and CSA values, or, alternatively, obtained by refining protein structures using these values as harmonic restraints in simulated annealing calculations. The Achillesā?? heel of this approach is the lack of suitable experiments for sequential assignment of the amide resonances. In this Article, we present a new pulse sequence that integrates proton driven spin diffusion (PDSD) with sensitivity-enhanced PISEMA in a 3D experiment ([1H,15N]-SE-PISEMA-PDSD). The incorporation of 2D 15N/15N spin diffusion experiments into this new 3D experiment leads to the complete and unambiguous assignment of the 15N resonances. The feasibility of this approach is demonstrated for the membrane protein sarcolipin reconstituted in magnetically aligned lipid bicelles. Taken with low electric field probe technology, this approach will propel the determination of sequential assignment as well as structure and topology of larger integral membrane proteins in aligned lipid bilayers.
Content Type Journal Article
Category Article
Pages 1-8
DOI 10.1007/s10858-011-9571-8
Authors
Kaustubh R. Mote, Department of Chemistry, University of Minnesota, Minneapolis, MN 55455-0431, USA
T. Gopinath, Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, 207 Pleasant St. S. E, Minneapolis, MN 55455, USA
Nathaniel J. Traaseth, Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, 207 Pleasant St. S. E, Minneapolis, MN 55455, USA
Jason Kitchen, National High Magnetic Field Laboratory, Tallahassee, FL 32310, USA
Peter L. Gorā??kov, National High Magnetic Field Laboratory, Tallahassee, FL 32310, USA
William W. Brey, National High Magnetic Field Laboratory, Tallahassee, FL 32310, USA
Gianluigi Veglia, Department of Chemistry, University of Minnesota, Minneapolis, MN 55455-0431, USA
(1)H assisted (13)C/(15)N heteronuclear correlation spectroscopy in oriented sample solid-state NMR of single crystal and magnetically aligned samples.
(1)H assisted (13)C/(15)N heteronuclear correlation spectroscopy in oriented sample solid-state NMR of single crystal and magnetically aligned samples.
(1)H assisted (13)C/(15)N heteronuclear correlation spectroscopy in oriented sample solid-state NMR of single crystal and magnetically aligned samples.
J Magn Reson. 2011 Apr 9;
Authors: Lin EC, Opella SJ
(1)H-irradiation under mismatched Hartmann-Hahn conditions provides an alternative mechanism for carrying out (15)N/(13)C transfers in triple-resonance heteronuclear correlation spectroscopy...
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1H Assisted 13C/15N Heteronuclear Correlation Spectroscopy in Oriented Sample Solid-State NMR of Single Crystal and Magnetically Aligned Samples
1H Assisted 13C/15N Heteronuclear Correlation Spectroscopy in Oriented Sample Solid-State NMR of Single Crystal and Magnetically Aligned Samples
Publication year: 2011
Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 9 April 2011</br>
Eugene C., Lin , Stanley J., Opella</br>
1H-irradiation under mismatched Hartmann-Hahn conditions provides an alternative mechanism for carrying out 15N/13C transfers in triple-resonance heteronuclear correlation spectroscopy (HETCOR) on stationary samples of single crystals and aligned samples of biopolymers, which...
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04-10-2011 12:52 PM
1H Assisted 13C/15N Heteronuclear Correlation Spectroscopy in Oriented Sample Solid-State NMR of Single Crystal and Magnetically Aligned Samples
1H Assisted 13C/15N Heteronuclear Correlation Spectroscopy in Oriented Sample Solid-State NMR of Single Crystal and Magnetically Aligned Samples
Publication year: 2011
Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 9 April 2011</br>
Eugene C., Lin , Stanley J., Opella</br>
1H-irradiation under mismatched Hartmann-Hahn conditions provides an alternative mechanism for carrying out 15N/13C transfers in triple-resonance heteronuclear correlation spectroscopy (HETCOR) on stationary samples of single crystals and aligned samples of biopolymers, which...
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Authors: Lorch M, Faham S, Kaiser C, Weber I, Mason AJ, Bowie JU, Glaubitz C
Several studies have demonstrated that it is viable to use microcrystalline preparations of water-soluble proteins as...
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[NMR paper] Solid-state NMR studies of magnetically aligned phospholipid membranes: taming lantha
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