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J Am Chem Soc. 2005 May 11;127(18):6893-901.

A suite of 2H NMR spin relaxation experiments for the measurement of RNA dynamics.

Author information

1
Protein Engineering Network Centers of Excellence and the Departments of Medical Genetics, Biochemistry and Chemistry, The University of Toronto, Toronto, Ontario, Canada, M5S 1A8.

Abstract

A suite of (2)H-based spin relaxation NMR experiments is presented for the measurement of molecular dynamics in a site-specific manner in uniformly (13)C, randomly fractionally deuterated ( approximately 50%) RNA molecules. The experiments quantify (2)H R(1) and R(2) relaxation rates that can subsequently be analyzed to obtain information about dynamics on a pico- to nanosecond time scale. Sensitivity permitting, the consistency of the data can be evaluated by measuring all five rates that are accessible for a spin 1 particle and establishing that the rates obey relations that are predicted from theory. The utility of the methodology is demonstrated with studies of the dynamics of a 14-mer RNA containing the UUCG tetraloop at temperatures of 25 and 5 degrees C. The high quality of the data, even at 5 degrees C, suggests that the experiments will be of use for the study of RNA molecules that are as large as 30 nucleotides.

PMID:
15869313
DOI:
10.1021/ja0427799
[Indexed for MEDLINE]

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