Squaramate-Modified Nucleotides and DNA for Specific Cross-Linking with Lysine-Containing Peptides and Proteins
Squaramate-Modified Nucleotides and DNA for Specific Cross-Linking with Lysine-Containing Peptides and Proteins
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DOI：
10.1002/anie.201906737
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发表时间：
2019-08-13
影响因子：
16.6
通讯作者：
Hocek, Michal
中科院分区：
文献类型：
--
作者：
Ivancova, Ivana;Pohl, Radek;Hocek, Michal
合成了方酸连接的 2'-脱氧胞苷 5'-O-三磷酸，发现它是修饰 DNA 的引物延伸或 PCR 合成中 KOD XL DNA 聚合酶的良好底物。所得的方酸酯连接的 DNA 与伯胺反应形成稳定的二酰胺键。该反应用于 DNA 与含 Cy5 和 Lys 的肽的生物缀合。方酸连接的 DNA 与组蛋白形成共价交联。这种反应性核苷酸具有核酸与胺、肽或蛋白质的其他生物缀合的潜力，无需任何外部试剂。
Squaramate-linked 2 '-deoxycytidine 5 '-O-triphosphate was synthesized and found to be good substrate for KOD XL DNA polymerase in primer extension or PCR synthesis of modified DNA. The resulting squaramate-linked DNA reacts with primary amines to form a stable diamide linkage. This reaction was used for bioconjugations of DNA with Cy5 and Lys-containing peptides. Squaramate-linked DNA formed covalent cross-links with histone proteins. This reactive nucleotide has potential for other bioconjugations of nucleic acids with amines, peptides or proteins without need of any external reagent.