title: Aqueous Heck cross-coupling preparation of acrylate-modified nucleotides and nucleoside triphosphates for polymerase synthesis of acrylate-labeled DNA.
abstract: Aqueous-phase Heck coupling methodology was developed for direct attachment of butyl acrylate to 5-iodoracil, 5-iodocytosine, 7-iodo-7-deazaadenine, and 7-iodo-7-deazaguanine 2'-deoxyribonucleoside 5'-O-monophosphates (dNMPs) and 5'-O-triphosphates (dNTPs) and compared with the classical approach of phosphorylation of the corresponding modified nucleosides. The 7-substituted 7-deazapurine nucleotides (dA(BA)MP, dA(BA)TP, dG(BA)MP, and dG(BA)TP) were prepared by the direct Heck coupling of nucleotides in good yields (35-55%), whereas the pyrimidine nucleotides reacted poorly and the corresponding BA-modified dNTPs were prepared by triphosphorylation of the modified nucleosides. The acrylate-modified dN(BA)TPs (N = A, C, and U) were good substrates for DNA polymerases and were used for enzymatic synthesis of acrylate-modified DNA by primer extension, whereas dG(BA)TP was an inhibitor of polymerases. The butyl acrylate group was found to be a useful redox label giving a strong reduction peak at -1.3 to -1.4 V in cyclic voltammetry.
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Gilead Sciences & IOCB Research Center , Flemingovo nám. 2, CZ-16610 Prague 6, Czech Republic.
authors.forename: Jitka
authors.initials: J
authors.lastname: Dadová
authors.forename: Pavlína
authors.initials: P
authors.lastname: Vidláková
authors.forename: Radek
authors.initials: R
authors.lastname: Pohl
authors.forename: Luděk
authors.initials: L
authors.lastname: Havran
authors.forename: Miroslav
authors.initials: M
authors.lastname: Fojta
authors.forename: Michal
authors.initials: M
authors.lastname: Hocek
mesh_terms: D000179:Acrylates / Q000138:chemical synthesis* / Q000737:chemistry*; D000225:Adenine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry* / Q000378:metabolism; D006147:Guanine / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000737:chemistry; D015394:Molecular Structure; D009705:Nucleosides / Q000138:chemical synthesis* / Q000737:chemistry; D009711:Nucleotides / Q000138:chemical synthesis* / Q000737:chemistry
chemical_list: C000589364:7-iodo-7-deazaadenine; D000179:Acrylates; D009705:Nucleosides; D009711:Nucleotides; D006147:Guanine; C032490:n-butyl acrylate; D004259:DNA-Directed DNA Polymerase; C036658:acrylic acid; D000225:Adenine