Longitudinal-relaxation-enhanced NMR experiments for the study of nucleic acids in solution.
Atomic-resolution information on the structure and dynamics of nucleic acids is essential for a better understanding of the mechanistic basis of many cellular processes. NMR spectroscopy is a powerful method for studying the structure and dynamics of nucleic acids; however, solution NMR studies are currently limited to relatively small nucleic acids at high concentrations. Thus, technological and methodological improvements that increase the experimental sensitivity and spectral resolution of NMR spectroscopy are required for studies of larger nucleic acids or protein-nucleic acid complexes. Here we introduce a series of imino-proton-detected NMR experiments that yield an over 2-fold increase in sensitivity compared to conventional pulse schemes. These methods can be applied to the detection of base pair interactions, RNA-ligand titration experiments, measurement of residual dipolar (15)N-(1)H couplings, and direct measurements of conformational transitions. These NMR experiments employ longitudinal spin relaxation enhancement techniques that have proven useful in protein NMR spectroscopy. The performance of these new experiments is demonstrated for a 10 kDa TAR-TAR*(GA) RNA kissing complex and a 26 kDa tRNA.
Farjon
Jonathan
J
Institut de Biologie Structurale-Jean-Pierre Ebel, UMR5075 CNRS-CEA-UJF, 41, rue Jules Horowitz, 38027 Grenoble Cedex, France.
Boisbouvier
Jérôme
J
Schanda
Paul
P
Pardi
Arthur
A
Simorre
Jean-Pierre
JP
Brutscher
Bernhard
B
D006678:HIV / Q000235:genetics; D006860:Hydrogen Bonding; D050111:Imino Sugars / Q000737:chemistry; D007700:Kinetics; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry*; D012366:RNA, Transfer, Val / Q000737:chemistry; D012367:RNA, Viral / Q000737:chemistry; D012996:Solutions
D050111:Imino Sugars; D012366:RNA, Transfer, Val; D012367:RNA, Viral; D012996:Solutions; D012313:RNA