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Assignment of the Nonexchanging Protons of the α ‐Spectrin SH3 Domain by Two‐ and Three‐ Dimensional 1 H– 13 C Solid‐State Magic‐Angle Spinning NMR and Comparison of Solution and Solid‐State Proton Chemical Shifts
Author(s) -
van Rossum BarthJan,
Castellani Federica,
Rehbein Kristina,
Pauli Jutta,
Oschkinat Hartmut
Publication year - 2001
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/1439-7633(20011203)2:12<906::aid-cbic906>3.0.co;2-m
Subject(s) - heteronuclear molecule , chemical shift , magic angle spinning , solid state nuclear magnetic resonance , chemistry , crystallography , homonuclear molecule , molecule , proton , microcrystalline , spectrin , nuclear magnetic resonance spectroscopy , nuclear magnetic resonance , analytical chemistry (journal) , stereochemistry , physics , organic chemistry , cell , biochemistry , quantum mechanics , cytoskeleton
The assignment of nonexchanging protons of a small microcrystalline protein, the α‐spectrin SH3 domain (7.2 kDa, 62 residues), was achieved by means of three‐dimensional (3D) heteronuclear ( 1 H– 13 C– 13 C) magic‐angle spinning (MAS) NMR dipolar correlation spectroscopy. With the favorable combination of a high B 0 ‐field, a moderately high spinning frequency, and frequency‐switched Lee‐Goldburg irradiation applied during 1 H evolution, a proton linewidth ≤0.5 ppm at 17.6 Tesla was achieved for the particular protein preparation used. A comparison of the solid‐state 1 H chemical shifts with the shifts found in solution shows a remarkable similarity, which reflects the identical protein structures in solution and in the solid. Significant differences between the MAS solid‐ and liquid‐state 1 H chemical shifts are only observed for residues that are located at the surface of the protein and that exhibit contacts between different SH3 molecules. In two cases, aromatic residues of neighboring SH3 molecules induce pronounced upfield ring‐current shifts for protons in the contact area.