Short, synthetic and selectively 13C-labeled RNA sequences for the NMR structure determination of protein-RNA complexes.
We report an optimized synthesis of all canonical 2'-O-TOM protected ribonucleoside phosphoramidites and solid supports containing [13C5]-labeled ribose moieties, their sequence-specific introduction into very short RNA sequences and their use for the structure determination of two protein-RNA complexes. These specifically labeled sequences facilitate RNA resonance assignments and are essential to assign a high number of sugar-sugar and intermolecular NOEs, which ultimately improve the precision and accuracy of the resulting structures. This labeling strategy is particularly useful for the study of protein-RNA complexes with single-stranded RNA in solution, which is rapidly an increasingly relevant research area in biology.
Wenter
Philipp
P
Institut des Science et Ingénierie Chimiques, Ecole Polytechnique Fédérale de Lausanne, EPFL-BCH, 1015 Lausanne, Switzerland.
Reymond
Luc
L
Auweter
Sigrid D
SD
Allain
Frédéric H-T
FH
Pitsch
Stefan
S
D002247:Carbon Isotopes; D007553:Isotope Labeling; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009843:Oligoribonucleotides / Q000138:chemical synthesis / Q000737:chemistry*; D009943:Organophosphorus Compounds / Q000737:chemistry; D038941:Polypyrimidine Tract-Binding Protein / Q000737:chemistry; D012313:RNA / Q000737:chemistry*; D016601:RNA-Binding Proteins / Q000737:chemistry*; D012263:Ribonucleosides / Q000737:chemistry; D012266:Ribose / Q000737:chemistry
D002247:Carbon Isotopes; D009843:Oligoribonucleotides; D009943:Organophosphorus Compounds; D016601:RNA-Binding Proteins; D012263:Ribonucleosides; C434331:phosphoramidite; D038941:Polypyrimidine Tract-Binding Protein; D012313:RNA; D012266:Ribose