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Solid‐state NMR study of the SH3 domain of α‐spectrin: application of 13 C– 15 N TEDOR and REDOR †
Author(s) -
Macholl Sven,
Sack Ingolf,
Limbach HansHeinrich,
Pauli Jutta,
Kelly Mark,
Buntkowsky Gerd
Publication year - 2000
Publication title -
magnetic resonance in chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.483
H-Index - 72
eISSN - 1097-458X
pISSN - 0749-1581
DOI - 10.1002/1097-458x(200007)38:7<596::aid-mrc713>3.0.co;2-k
Subject(s) - chemistry , solid state nuclear magnetic resonance , magic angle spinning , nuclear magnetic resonance spectroscopy , nuclear magnetic resonance , residual dipolar coupling , spectrin , magnetic dipole–dipole interaction , spectroscopy , alanine , dipole , deuterium , residue (chemistry) , analytical chemistry (journal) , crystallography , stereochemistry , atomic physics , amino acid , physics , chromatography , biochemistry , cytoskeleton , cell , organic chemistry , quantum mechanics
A fully 13 C– 15 N‐labeled and a selectively alanine‐ 13 C β tryptophan‐ 15 N ring ‐labeled sample of the Src homology region 3 (SH3) domain of α‐spectrin (chicken), a 62 residue protein, were biosynthesized and studied by solid‐state cross‐polarization magic angle spinning (CP/MAS) NMR, 13 C– 15 N rotational echo double resonance (REDOR) and 15 N– 13 C transferred echo double resonance (TEDOR) spectroscopy. In the first part of the study it is shown that spectral editing with the TEDOR sequence leads to a drastic simplification of the 13 C MAS spectrum of the fully labeled sample, allowing the resolved spectroscopy of groups of 13 C nuclei, according to their distance to neighboring 15 N nuclei. In the second part of the study the inter‐residual distance between the alanine residue Ala55 and the tryptophan residue Trp42 was determined by the measurement of the dipolar coupling between Ala‐ 13 C β and Trp‐ 15 N ring , yielding a dipolar coupling of 48 ± 8 Hz, which after correction for fast molecular vibrations gives a value of 53 ± 8 Hz, corresponding to a CN distance of 3.85 ± 0.25 Hz. The result is compared to the CN distances obtained by x‐ray diffraction and liquid‐state NMR. Copyright © 2000 John Wiley & Sons, Ltd.