Combinatorial synthesis of thrombin-binding aptamers containing iso-guanine.
A library of all possible substitutions of guanine by iso-guanine (iG) in the thrombin aptamer was prepared by split and mix synthesis. A colorimetric assay was used to screen for functional oligomers in the library. Colorimetrically active oligonucleotides were selected and sequenced by the Maxam-Gilbert method. The sequenced oligonucleotides were individually resynthesized, and their affinities for thrombin were assayed by isothermal titration calorimetry. Three aptamer sequences containing iG were found to have enhanced binding activity to human alpha-thrombin compared to the parent aptamer.
Nallagatla
Subba Rao
SR
Department of Chemistry, University of California, Riverside, California 92521, USA.
Heuberger
Benjamin
B
Haque
Azizul
A
Switzer
Christopher
C
D052157:Aptamers, Nucleotide / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D002151:Calorimetry; D003124:Colorimetry; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D006147:Guanine / Q000138:chemical synthesis / Q000737:chemistry*; D006801:Humans; D007536:Isomerism; D008958:Models, Molecular; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D013917:Thrombin / Q000378:metabolism*; D017186:Titrimetry
D052157:Aptamers, Nucleotide; D009841:Oligonucleotides; C081676:thrombin aptamer; D006147:Guanine; D013917:Thrombin