Synthesis of Fluorophosphate Nucleotide Analogs and Their Characterization as Tools for 19F NMR Studies.
Synthesis of Fluorophosphate Nucleotide Analogs and Their Characterization as Tools for 19F NMR Studies.
J Org Chem. 2015 Mar 27;
Authors: Baranowski MR, Nowicka A, Rydzik AM, Warminski M, Kasprzyk R, Wojtczak BA, Wojcik J, Claridge TD, Kowalska J, Jemielity J
Abstract
To broaden the scope of existing methods based on 19F nucleotide labeling, we developed a new method for the synthesis of fluorophosphate (oligo)nucleotide analogs containing an O to F substitution at the terminal position of the (oligo)phosphate moiety and evaluated them as tools for 19F NMR studies. Using three efficient and comprehensive synthetic approaches based on phosphorimidazolide chemistry and tetra-n-butylammonium fluoride, fluoromonophosphate or fluorophosphate imidazolide as fluorine sources, we prepared over 30 fluorophosphate-containing nucleotides, varying in nucleobase type (A, G, C, U, m7G), phosphate chain length (from mono- to tetra-) and presence of additional phosphate modifications (thio-, borano-, imido-, methylene-). Using fluorophosphate imidazolide as fluorophosphorylating reagent for 5'-phosphorylated oligos we also synthesized oligonucleotide 5'-(2-fluorodiphosphates), which are potentially useful as 19F NMR hybridization probes. The compounds were characterized by 19F NMR, and evaluated as 19F NMR molecular probes. We found that fluorophosphate nucleotide analogs can be used to monitor activity of enzymes with various specificities and metal-ion requirements, including human DcpS enzyme - a therapeutic target for spinal muscular atrophy. The compounds can also serve as reporter ligands for protein binding studies, as exemplified by studying interaction of fluorophosphate mRNA cap analogs with eukaryotic translation initiation factor (eIF4E).
PMID: 25816092 [PubMed - as supplied by publisher]
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