Lincoln Scott - New RNA labeling methods offer dramatic sensitivity enhancements in 2H NMR relaxation spectra.

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  J Am Chem   Soc. 2006 Jul 26;128(29):9346-7.


  A new labeling strategy is presented that greatly facilitates the   measurement of 2H spin relaxation rates in RNA molecules as a   probe of pico- to nanosecond time scale dynamics. In this   labeling scheme the sugar positions are uniformly 13C-labeled,   with position 2' protonated and all other sites on the sugar   deuterated. Pulse sequences are presented for measurement of 2H   R1 and R2 relaxation rates at positions 1', 3', and 4' with   sensitivity gains that are on the order of 5-fold relative to   previous methods that employed random fractional deuteration. The   improved sensitivity is transformative and facilitates the study   of motion in moderately sized RNA molecules with good   sensitivity. The utility of the approach is demonstrated with an   application to HIV-2 TAR, where the site-specific measures of   molecular dynamics at sugar positions obtained here complement   previous studies of dynamics at aromatic sites in the molecule.

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