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Characterization of G‐Quadruplex/Hairpin Transitions of RNAs by 19 F NMR Spectroscopy
Author(s) -
Granqvist Lotta,
Virta Pasi
Publication year - 2016
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201602898
Subject(s) - guanosine , chemistry , rna , nuclear magnetic resonance spectroscopy , crystallography , cytidine , nmr spectra database , two dimensional nuclear magnetic resonance spectroscopy , phosphorus 31 nmr spectroscopy , phosphoramidite , oligonucleotide , stereochemistry , spectral line , organic chemistry , biochemistry , dna , enzyme , physics , astronomy , gene
2′‐ O ‐[(4‐Trifluoromethyl‐triazol‐1‐yl)methyl] reporter groups have been incorporated into guanosine‐rich RNA models (including a known bistable Qd/Hp RNA and two G‐rich regions of mRNA of human prion protein, PrP) and applied for the 19 F NMR spectroscopic characterization of plausible G‐quadruplex/hairpin (Qd/Hp) transitions in these RNA structures. For the synthesis of the CF 3 ‐labeled RNAs, phosphoramidite building blocks of 2′‐ O ‐[(4‐CF 3 ‐triazol‐1‐yl)methyl] nucleosides (cytidine, adenosine, and guanosine) were prepared and used as an integral part of the standard solid‐phase RNA synthesis. The obtained 19 F NMR spectra supported the usual characterization data (obtained by UV‐ and CD‐melting profiles and by 1 H NMR spectra of the imino regions) and additionally gave more detailed information on the Qd/Hp transitions. The molar fractions of the secondary structural species (Qd, Hp) upon thermal denaturation and under varying ionic conditions could be determined from the intensities and shifts of the 19 F NMR signals. For a well‐behaved Qd/Hp transition, thermodynamic parameters could be extracted.