Isotope labeling and segmental labeling of larger RNAs for NMR structural studies.
NMR spectroscopy has become substantial in the elucidation of RNA structures and their complexes with other nucleic acids, proteins or small molecules. Almost half of the RNA structures deposited in the Protein Data Bank were determined by NMR spectroscopy, whereas NMR accounts for only 11% for proteins. Recent improvements in isotope labeling of RNA have strongly contributed to the high impact of NMR in RNA structure determination. In this book chapter, we review the advances in isotope labeling of RNA focusing on larger RNAs. We start by discussing several ways for the production and purification of large quantities of pure isotope labeled RNA. We continue by reviewing different strategies for selective deuteration of nucleotides. Finally, we present a comparison of several approaches for segmental isotope labeling of RNA. Selective deuteration of nucleotides in combination with segmental isotope labeling is paving the path for studying RNAs of ever increasing size.
Swiss Federal Institute of Technology Zürich, Zürich, Switzerland.
Olivier
O
Duss
Peter J
PJ
Lukavsky
Frédéric H-T
FH
Allain
D007553:Isotope Labeling / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry*
D012313:RNA