DNP enhanced frequency-selective TEDOR experiments in bacteriorhodopsin |
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Authors: | Vikram S. Bajaj Melody L. Mak-Jurkauskas Marina Belenky Judith Herzfeld Robert G. Griffin |
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Affiliation: | 1. Francis Bitter Magnet Laboratory, Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States;2. Department of Chemistry, Brandeis University, Waltham, MA 02454, United States |
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Abstract: | We describe a new approach to multiple 13C–15N distance measurements in uniformly labeled solids, frequency-selective (FS) TEDOR. The method shares features with FS-REDOR and ZF- and BASE-TEDOR, which also provide quantitative 15N–13C spectral assignments and distance measurements in U-[13C,15N] samples. To demonstrate the validity of the FS-TEDOR sequence, we measured distances in [U-13C,15N]-asparagine which are in good agreement with other methods. In addition, we integrate high frequency dynamic nuclear polarization (DNP) into the experimental protocol and use FS-TEDOR to record a resolved correlation spectrum of the Arg-13Cγ–15Nε region in [U-13C,15N]-bacteriorhodopsin. We resolve six of the seven cross-peaks expected based on the primary sequence of this membrane protein. |
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