Complementary segmental labeling of large RNAs: economic preparation and simplified NMR spectra for measurement of more RDCs |
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Authors: | Tzakos Andreas G Easton Laura E Lukavsky Peter J |
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Institution: | MRC, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom. |
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Abstract: | NMR structure determination of large RNAs is often restricted by limited RDC information caused by chemical shift degeneracy. We established a general, time- and cost-effective methodology for the preparation of 13C/15N complementary labeled RNAs from a single plasmid. Applying this method to the 25 kDa BC1-DTE RNA, we were able to resolve severe chemical shift degeneracy, thereby almost doubling the number of RDC restraints in comparison to the conventional 13C,15N uniform-labeled RNA. |
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