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Q Rev Biophys. 2015 Feb;48(1):35-116. doi: 10.1017/S0033583514000122.

Visualizing transient dark states by NMR spectroscopy.

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1
Laboratory of Chemical Physics,National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health,Bethesda,Maryland 20892-0520,USA.

Abstract

Myriad biological processes proceed through states that defy characterization by conventional atomic-resolution structural biological methods. The invisibility of these 'dark' states can arise from their transient nature, low equilibrium population, large molecular weight, and/or heterogeneity. Although they are invisible, these dark states underlie a range of processes, acting as encounter complexes between proteins and as intermediates in protein folding and aggregation. New methods have made these states accessible to high-resolution analysis by nuclear magnetic resonance (NMR) spectroscopy, as long as the dark state is in dynamic equilibrium with an NMR-visible species. These methods - paramagnetic NMR, relaxation dispersion, saturation transfer, lifetime line broadening, and hydrogen exchange - allow the exploration of otherwise invisible states in exchange with a visible species over a range of timescales, each taking advantage of some unique property of the dark state to amplify its effect on a particular NMR observable. In this review, we introduce these methods and explore two specific techniques - paramagnetic relaxation enhancement and dark state exchange saturation transfer - in greater detail.

PMID:
25710841
DOI:
10.1017/S0033583514000122
[Indexed for MEDLINE]
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