148692-01-7Relevant articles and documents
CCR5 RNA Pseudoknots: Residue and Site-Specific Labeling correlate Internal Motions with microRNA Binding
Chen, Bin,Longhini, Andrew P.,Nu?baumer, Felix,Kreutz, Christoph,Dinman, Jonathan D.,Dayie, T. Kwaku
, p. 5462 - 5468 (2018)
Conformational dynamics of RNA molecules play a critical role in governing their biological functions. Measurements of RNA dynamic behavior sheds important light on sites that interact with their binding partners or cellular stimulators. However, such measurements using solution-state NMR are difficult for large RNA molecules (>70 nt; nt=nucleotides) owing to severe spectral overlap, homonuclear 13C scalar couplings, and line broadening. Herein, a strategic combination of solid-phase synthesis, site-specific isotopic labeled phosphoramidites, and enzymatic ligation is introduced. This approach allowed the position-specific insertion of isotopic probes into a 96 nt CCR5 RNA fragment. Accurate measurements of functional dynamics using the Carr–Purcell–Meiboom–Gill (CPMG) relaxation dispersion (RD) experiments enabled extraction of the exchange rates and populations of this RNA. NMR chemical shift perturbation analysis of the RNA/microRNA-1224 complex indicated that A90-C1′ of the pseudoknot exhibits similar changes in chemical shift observed in the excited state. This work demonstrates the general applicability of a NMR-labeling strategy to probe functional RNA structural dynamics.
Regioselective protection of the 2'-hydroxyl group of N-acyl-3',5'-O-di(t-butyl)silanediylnucleoside derivatives by use of t-BuMgCl and 2-(trimethylsilyl)ethoxymethyl chloride
Wada,Tobe,Nagayama,Furusawa,Sekine
, p. 1683 - 1684 (2007/10/02)
Regioselective 2'-O-protection of N-protected 3',5'-O-di(t-butyl)silanediylribonucleoside derivatives with the 2-(trimethylsilyl)ethoxymethyl (SEM) group has been achieved by use of t-BuMgCl and 2-(trimethylsilyl)ethoxymethyl chloride. The former was used