Novel monofunctional substrates of polynucleotide phosphorylase.  The "single-addition" of 2'(3')-O-dihydrocinnamoyl-nucleoside 5'-diphosphate to a primer oligonucleotide.
A method was developed for stepwise wynthesis of oligonucleotides of difined wequence using 2'(3')-O-dihydrocinnamoyl-nucleoside 5'-diphosphates as substrates for polynucleotide phosphorylase [ED 2.7.7.8].  Polynucleotide phosphorylase from Thermus thermophilus catalyzed the transfer of one 2'(3')-blocked ADP to the 3'-terminus of the primer trinucleoside diphosphate, ApApA.  The product was 2'(3')-substituted triadenylyladenosine.  The blocking group, dihydrocinnamoyl, could be removed completely from the product without destruction of the phosphodiester bond using alpha-chymotrypsin [ED 3.4.21.1] at neutral pH.
D001419:Bacteria / Q000201:enzymology; D002934:Cinnamates; D004355:Drug Stability; D006358:Hot Temperature; D007700:Kinetics; D009841:Oligonucleotides / Q000138:chemical synthesis; D011117:Polyribonucleotide Nucleotidyltransferase / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013997:Time Factors
D002934:Cinnamates; D009841:Oligonucleotides; D011117:Polyribonucleotide Nucleotidyltransferase