Abstract:
Glyoxal-linked 2\'-deoxyuridine 5\'-O-mono- and triphosphates were synthesized through a CuAAC click reaction of 4-azidophenylglyoxal or a Sonogashira reaction of 4-bromophenylglyoxal with 5-ethynyl-dUMP or -dUTP. The triphosphates were used as substrates for enzymatic synthesis of modified DNA probes with KOD XL DNA polymerase. The glyoxal-linked nucleotides reacted with arginine-containing peptides to form stable imizadolone-linked conjugates. This reactive glyoxal modification in DNA was used for efficient bioconjugations and crosslinking with Arg-containing peptides or proteins (e. g., histones) and was found to be more reactive than previously reported 1,3-diketone-linked DNA probes.
摘要:
乙二醛连接的2'-脱氧尿苷5'-O-单磷酸和三磷酸是通过4-叠氮苯基乙二醛的CuAAC点击反应或4-溴苯乙二醛与5-乙炔基-dUMP或-dUTP的Sonogashira反应合成的。三磷酸盐用作底物,用于用KODXLDNA聚合酶酶促合成修饰的DNA探针。乙二醛连接的核苷酸与含精氨酸的肽反应以形成稳定的伊米唑酮连接的缀合物。DNA中的这种反应性乙二醛修饰用于与含Arg的肽或蛋白质进行有效的生物缀合和交联(例如Procedure,组蛋白),并且发现比先前报道的1,3-二酮连接的DNA探针更具反应性。