Solution NMR Experiment for Measurement of 15N-1H Residual Dipolar Couplings in Large Proteins and Supramolecular Complexes.
 

Solution NMR Experiment for Measurement of 15N-1H Residual Dipolar Couplings in Large Proteins and Supramolecular Complexes.

Related Articles Solution NMR Experiment for Measurement of 15N-1H Residual Dipolar Couplings in Large Proteins and Supramolecular Complexes.

J Am Chem Soc. 2015 Aug 21;

Authors: Eletsky A, Pulavarti SV, Beaumont V, Gollnick P, Szyperski T

Abstract
NMR residual dipolar couplings (RDCs) are exquisite probes of protein structure and dynamics. A new solution NMR experiment named 2D SE2 J-TROSY is presented to measure N-H RDCs for proteins and supramolecular complexes in excess of 200 kDa. This enables validation and refinement of their X-ray crystal and solution NMR structures, and the characterization of structural and dynamic changes occurring upon complex formation. Accurate N-H RDCs were measured at 750 MHz 1H resonance frequency for 11-mer 93 kDa 2H,15N-labeled Trp RNA-binding Attenuator Protein (TRAP) tumbling with a correlation time ?c of 120 ns. This is about twice as long as for the most slowly tumbling system, for which N-H RDCs could be measured so far, and corresponds to molecular weights ~200 kDa at 25 oC. Fur-thermore, due to the robustness of SE2 J-TROSY with respect to residual 1H density from exchangeable protons, increased sensitivity at 1H resonance frequencies around 1 GHz promis-es to enable N-H RDC measurement for even larger systems.


PMID: 26293598 [PubMed - as supplied by publisher]



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