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Default An enhanced sensitivity methyl 1 H triple-quantum pulse scheme for measuring diffusion constants of macromolecules

An enhanced sensitivity methyl 1 H triple-quantum pulse scheme for measuring diffusion constants of macromolecules

Abstract

We present a pulse scheme that exploits methyl 1H triple-quantum (TQ) coherences for the measurement of diffusion rates of slowly diffusing molecules in solution. It is based on the well-known stimulated echo experiment, with encoding and decoding of TQ coherences. The size of quantifiable diffusion coefficients is thus lowered by an order of magnitude with respect to single-quantum (SQ) approaches. Notably, the sensitivity of the scheme is high, approximately ¬ĺ that of the corresponding single quantum experiment, neglecting relaxation losses, and on the order of a factor of 4 more sensitive than a previously published sequence for AX3 spin systems (Zheng et al. in JMR 198:271‚??274, 2009) for molecules that are only 13C labeled at the methyl carbon position. Diffusion coefficients measured from TQ- and SQ-based experiments recorded on a range of protein samples are in excellent agreement. We present an application of this technique to the study of phase-separated proteins where protein concentrations in the condensed phase can exceed 400¬*mg/mL, diffusion coefficients can be as low as ~10‚??9 cm2s‚??1 and traditional SQ experiments fail.



Source: Journal of Biomolecular NMR
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