A suite of 2H NMR spin relaxation experiments for the measurement of RNA dynamics.
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.
Vallurupalli
Pramodh
P
Protein Engineering Network Centers of Excellence and the Departments of Medical Genetics, Biochemistry and Chemistry, The University of Toronto, Toronto, Ontario, Canada, M5S 1A8.
Kay
Lewis E
LE
D002247:Carbon Isotopes; D003903:Deuterium; D006147:Guanine / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D012313:RNA / Q000737:chemistry*; D013696:Temperature; D013816:Thermodynamics; D014498:Uracil / Q000737:chemistry
D002247:Carbon Isotopes; D014498:Uracil; D006147:Guanine; D012313:RNA; D003903:Deuterium