Abstract：
Reactive N-hydroxy-9-azabicyclo[3.3.1]nonane (ABNOH) linked 2\'-deoxyuridine 5\'-O-mono- and triphosphates were synthesized through a CuAAC click reaction of ABNO-H-PEG4-N3 with 5-ethynyl-dUMP or -dUTP. The modified triphosphate was used as substrate for enzymatic synthesis of modified DNA probes with KOD XL DNA polymerase. The keto-ABNO radical reacted with tryptophan (Trp) and Trp-containing peptides to form a stable 3-fused hexahydropyrrolo-indole conjugates. Similarly modified ABNO-H-linked nucleotides reacted with Trp-containing peptides to form a stable conjugate in the presence but surprisingly even in the absence of NaNO2 (presumably through activation by O2). The reactive ABNO-H-modified DNA probe was used for bioconjugations and crosslinking with Trp-containing peptides or proteins.
摘要：
通过ABNO-H-PEG4-N3与5-乙炔基-dUMP或-dUTP的CuAAC点击反应合成反应性N-羟基-9-氮杂双环[3.3.1]壬烷（ABNOH）连接的2'-脱氧尿苷5'-O-单-和三磷酸盐。修饰的三磷酸酯用作底物，用于用KODXLDNA聚合酶酶促合成修饰的DNA探针。酮-ABNO自由基与色氨酸（Trp）和含Trp的肽反应，形成稳定的3-融合六氢吡咯并吲哚缀合物。类似地,修饰的ABNO-H-连接的核苷酸与含Trp的肽反应以在存在下形成稳定的缀合物,但令人惊讶地甚至在不存在NaNO2的情况下(推测通过O2活化)。反应性ABNO-H修饰的DNA探针用于与含Trp的肽或蛋白质的生物缀合和交联。
