caa a22 4500 445325275 CHVBK 20180317142718.0 cr unu---uuuuu 170323e20111101xx s 000 0 eng 10.1007/s10858-011-9571-8 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9571-8 Multidimensional oriented solid-state NMR experiments enable the sequential assignment of uniformly 15N labeled integral membrane proteins in magnetically aligned lipid bilayers [Elektronische Daten] [Kaustubh Mote, T. Gopinath, Nathaniel Traaseth, Jason Kitchen, Peter Gor'kov, William Brey, Gianluigi Veglia] 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 Achilles' 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. Springer Science+Business Media B.V., 2011 Oriented solid-state NMR (OSS - NMR) nationallicence Membrane proteins nationallicence Sequential assignment nationallicence Sarcolipin nationallicence Magnetically aligned bicelles nationallicence Proton driven spin diffusion nationallicence PISEMA nationallicence Sensitivity-enhancement nationallicence SLN : Sarcolipin nationallicence SERCA : sarcoplasmic reticulum Ca2+-ATPase nationallicence DMPC : 1,2-dimyristoyl-sn-glycero-3-phosphocholine nationallicence D6PC : 1,2-dihexanoyl-sn-glycero-3-phosphocholine nationallicence POPC : 1-palmitoyl,2-oleyl-sn-glycero-3-phosphocholine nationallicence SE-PISEMA : Sensitivity enhanced polarization inversion spin exchange at magic angle nationallicence PISA : Polar index slant angle nationallicence Mote Kaustubh Department of Chemistry, University of Minnesota, 55455-0431, Minneapolis, MN, USA aut Gopinath T. Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, 207 Pleasant St. S. E, 55455, Minneapolis, MN, USA aut Traaseth Nathaniel Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, 207 Pleasant St. S. E, 55455, Minneapolis, MN, USA aut Kitchen Jason National High Magnetic Field Laboratory, 32310, Tallahassee, FL, USA aut Gor'kov Peter National High Magnetic Field Laboratory, 32310, Tallahassee, FL, USA aut Brey William National High Magnetic Field Laboratory, 32310, Tallahassee, FL, USA aut Veglia Gianluigi Department of Chemistry, University of Minnesota, 55455-0431, Minneapolis, MN, USA aut Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 339-346 0925-2738 51:3<339 2011 51 10858 https://doi.org/10.1007/s10858-011-9571-8 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9571-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mote Kaustubh Department of Chemistry, University of Minnesota, 55455-0431, Minneapolis, MN, USA aut NATIONALLICENCE 700 1- Gopinath T. Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, 207 Pleasant St. S. E, 55455, Minneapolis, MN, USA aut NATIONALLICENCE 700 1- Traaseth Nathaniel Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, 207 Pleasant St. S. E, 55455, Minneapolis, MN, USA aut NATIONALLICENCE 700 1- Kitchen Jason National High Magnetic Field Laboratory, 32310, Tallahassee, FL, USA aut NATIONALLICENCE 700 1- Gor'kov Peter National High Magnetic Field Laboratory, 32310, Tallahassee, FL, USA aut NATIONALLICENCE 700 1- Brey William National High Magnetic Field Laboratory, 32310, Tallahassee, FL, USA aut NATIONALLICENCE 700 1- Veglia Gianluigi Department of Chemistry, University of Minnesota, 55455-0431, Minneapolis, MN, USA aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 339-346 0925-2738 51:3<339 2011 51 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer