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3D C(CC)H TOCSY Experiment for Assigning Protons and Carbons in Uniformly13C- and Selectively2H-Labeled RNA

✍ Scribed by Kwaku T. Dayie; Thomas J. Tolbert; James R. Williamson

Publisher: Elsevier Science
Year: 1998
Tongue: English
Weight: 134 KB
Volume: 130
Category: Article
ISSN: 1090-7807
DOI: 10.1006/jmre.1997.1286

No coin nor oath required. For personal study only.

✦ Synopsis

We have prepared RNAs uniformly 13C-labeled in the ribose ring and with the H3',H4',H5'/H5" protons specifically replaced with deuterons (3',4',5'/5"-2H-13C, termed d4-13C ribose), which offers the advantage of spectral simplification without sacrifice of sensitivity for the remaining protons. A 3D C(CC)H TOCSY experiment that uses a combination of cross-polarization transfer and deuterium decoupling improves the transfer of magnetization around the ribose ring and facilitates assignment of all ribose carbons and H1' and H2' protons in a 30-nucleotide RNA from HIV-2. These and other combinations of labeling and pulse sequence methodology should prove invaluable for the study of large RNAs, RNA-ligand, and RNA-protein complexes.

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