Efficient synthesis of [2'-18O]uridine and its incorporation into oligonucleotides: a new tool for mechanistic study of nucleotidyl transfer reactions by isotope effect analysis.
Lack of sufficient quantities of isotopically labeled materials has precluded the use of heavy atom isotope effects to investigate mechanisms of nucleotidyl transfer reactions in nucleic acids. Here we achieve regioselective opening of 2,2'-cyclouridine with [(18)O2]benzoic acid/potassium hydride, allowing an efficient "one-pot" synthesis of [2'-18O]uridine in 88% yield. Conversion to the corresponding phosphoramidite enables solid-phase synthesis of [2'-(18)O] RNA substrates for isotope effect studies with nucleotidyl transferases and hydrolases.
Department of Biochemistry & Molecular Biology, Department of Chemistry, and Howard Hughes Medical Institute, The University of Chicago, MC 1028, Chicago, Illinois 60637, USA.
Qing
Q
Dai
John K
JK
Frederiksen
Vernon E
VE
Anderson
Michael E
ME
Harris
Joseph A
JA
Piccirilli
D009690:Nucleic Acid Conformation; D009711:Nucleotides / Q000737:chemistry*; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry*; D010103:Oxygen Isotopes; D013237:Stereoisomerism; D014529:Uridine / Q000138:chemical synthesis* / Q000737:chemistry
D009711:Nucleotides; D009841:Oligonucleotides; D010103:Oxygen Isotopes; D014529:Uridine