title: ¹⁹F-Site-Specific-Labeled Nucleotides for Nucleic Acid Structural Analysis by NMR.
abstract: Naturally occurring RNA lacks fluorine-19 ((19)F), thus, their specifically fluorinated counterparts are particularly well suited to noninvasively monitoring the dynamic conformational properties and ligand-binding interactions of the RNA. For nuclear magnetic resonance (NMR) spectroscopy, (19)F-NMR of fluorine-substituted RNA provides an attractive, site-specific probe for structure determination in solution. Advantages of (19)F include high NMR sensitivity (83% of (1)H), high natural abundance (100%), and the extreme sensitivity of (19)F to the chemical environment leading to a large range of chemical shifts. The preparation of base-substituted 2-fluoropurine and 5-fluoropyrimidine 5'-triphosphates (2F-ATP/5F-CTP/5F-UTP) can be carried out using efficient enzymatic synthesis methods. Both pyrimidine analogs, 5-fluorouridine and 5-fluorocytidine, as well as, 2-fluoroadenosine are readily incorporated into RNA transcribed in vitro using T7 RNA polymerase.
authors.affiliation: Cassia LLC, San Diego, California, USA.
authors.forename: Lincoln G
authors.initials: LG
authors.lastname: Scott
authors.affiliation: Nutrition Research Institute, University of North Carolina at Chapel Hill, Kannapolis, North Carolina, USA. Electronic address: mirkohennig@yahoo.com.
authors.forename: Mirko
authors.initials: M
authors.lastname: Hennig
mesh_terms: D005461:Fluorine / Q000737:chemistry; D066235:Fluorine-19 Magnetic Resonance Imaging / Q000379:methods*; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009690:Nucleic Acid Conformation; D009711:Nucleotides / Q000737:chemistry; D012313:RNA / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry
chemical_list: D008024:Ligands; D009711:Nucleotides; D012996:Solutions; D005461:Fluorine; D012313:RNA
keywords: 2F-ATP; 5F-CTP; 5F-UTP; Enzymatic synthesis; Fluorine-19 NMR; HIV TAR; Ligand binding; Residual dipolar coupling; Solvent-induced isotope shift