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• W4200475268 abstract "The concentrations of specific macromolecular species can be quantified using diagnostic tools that rely on molecular recognition by nucleic acid aptamers. One such approach involves the formation of osmium tetroxide 2,2'-bipyridine protein adducts, followed by electrochemical detection of analytes that bind specifically to electrode-tethered aptamers. In conjunction with a 27-mer DNA aptamer that binds specifically to exosite II on human alpha thrombin, this technique permits, in theory, a highly sensitive diagnostic tool for the quantification of serum thrombin levels. However, thrombin's aptamer binding site is lined by two tryptophan residues and the conjugation of bulky osmium groups to these residues weakens aptamer binding by an estimated 4 to 12 kcal/mol, undermining detection sensitivity. Therefore, we have rationally modified this DNA aptamer to strengthen its thrombin binding in the presence of conjugated osmium. Specifically, aptamers carrying long hydrophobic thymine derivatives in place of guanine 21 have binding affinities for osmium-conjugated thrombin that are enhanced by 10 to 15 kcal/mol, suggesting that these modified aptamers may be effective in a highly sensitive electrochemical sensor for the quantification of low concentrations of thrombin. Our approach of using molecular simulation to subtly re-engineer a DNA aptamer may be generally applicable for the optimization of other macromolecular binding interfaces.Communicated by Ramaswamy H. Sarma" @default.
• W4200475268 created "2021-12-31" @default.
• W4200475268 creator A5018256607 @default.
• W4200475268 creator A5033674195 @default.
• W4200475268 date "2021-12-11" @default.
• W4200475268 modified "2023-09-25" @default.
• W4200475268 title "Design of a protein-targeted DNA aptamer using atomistic simulation" @default.
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• W4200475268 doi "https://doi.org/10.1080/07391102.2021.2011414" @default.
• W4200475268 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/34895068" @default.
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