Backbone and side chain assignment strategies for multiply labeled membrane peptides and proteins in the solid state |
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Authors: | Petkova Aneta T Baldus Marc Belenky Marina Hong Mei Griffin Robert G Herzfeld Judith |
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Institution: | Department of Chemistry and Keck Institute for Cellular Visualization, Brandeis University, 02454-9110, Waltham, MA, USA. |
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Abstract: | We demonstrate that the SPECIFIC CP technique can be used to obtain heteronuclear correlation (HETCOR) spectra of peptide backbones with greater efficiency than conventional HETCOR methods. We show that similar design principles can be employed to achieve selective homonuclear polarization transfer mediated through dipolar or scalar couplings. Both approaches are demonstrated in a tripeptide with uniform 15N and 13C labeling, and with uniform 15N labeling and natural abundance 13C. In other applications, the high efficiency of the heteronuclear SPECIFIC CP transfer allows discrimination of single amide signals in the 248-residue membrane protein bacteriorhodopsin (bR). In particular, variations are detected in the ordering of the Ala81-Arg82 peptide bond among the photocycle intermediates of bR and SPECIFIC CP is used to correlate 15N and 13C signals from the three Val-Pro peptide bonds. |
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