American Chemical Society
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Accurate Determination of Order Parameters from 1H,15N Dipolar Couplings in MAS Solid-State NMR Experiments

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journal contribution
posted on 2009-10-07, 00:00 authored by Veniamin Chevelkov, Uwe Fink, Bernd Reif
A reliable site-specific estimate of the individual N−H bond lengths in the protein backbone is the fundamental basis of any relaxation experiment in solution and in the solid-state NMR. The N−H bond length can in principle be influenced by hydrogen bonding, which would result in an increased N−H distance. At the same time, dynamics in the backbone induces a reduction of the experimental dipolar coupling due to motional averaging. We present a 3D dipolar recoupling experiment in which the 1H,15N dipolar coupling is reintroduced in the indirect dimension using phase-inverted CP to eliminate effects from rf inhomogeneity. We find no variation of the N−H dipolar coupling as a function of hydrogen bonding. Instead, variations in the 1H,15N dipolar coupling seem to be due to dynamics of the protein backbone. This is supported by the observed correlation between the HN−N dipolar coupling and the amide proton chemical shift. The experiment is demonstrated for a perdeuterated sample of the α-spectrin SH3 domain. Perdeuteration is a prerequisite to achieve high accuracy. The average error in the analysis of the H−N dipolar couplings is on the order of ±370 Hz (±0.012 Å) and can be as small as 150 Hz, corresponding to a variation of the bond length of ±0.005 Å.