寡核苷酸的血红素衍生物定向修饰的动力学研究

Kinetic study of the addressed modification by hemin derivatives of oligonucleotides.

作者信息

Frolova E I, Fedorova O S, Knorre D G

机构信息

Institute of Bioorganic Chemistry, Siberian Division of Russian Academy of Sciences, Novosibirsk.

出版信息

Biochimie. 1993;75(1-2):5-11. doi: 10.1016/0300-9084(93)90018-n.

DOI:10.1016/0300-9084(93)90018-n

PMID:8504178

Abstract

Kinetics of oligonucleotide pd(TGAATGGGAAGA) modification by a hemin derivative of the complementary oligonucleotide pd(TTCCCATT) in the presence of hydrogen peroxide was investigated. The treatment of experimental data permitted to evaluate the association and rate constants at 25 degrees C: Kx = (3.40 +/- 0.38) x 10(5) M-1 (association constant of the reagent with the target), kd = 152 +/- 6 M-1 min-1 (degradation constant of the hemin group of the reagent in a parallel reaction), ko = 51.0 +/- 1.7 M-1 min-1 (target modification constant in the reactive duplex). The modification of DNA is incomplete due to competition of the modification reaction with the degradation of the hemin group of the reagent in a parallel reaction.

摘要

研究了在过氧化氢存在下，互补寡核苷酸pd(TTCCCATT)的血红素衍生物对寡核苷酸pd(TGAATGGGAAGA)的修饰动力学。对实验数据的处理使得能够评估25℃时的缔合常数和速率常数：Kx = (3.40 ± 0.38) x 10(5) M-1（试剂与靶标的缔合常数），kd = 152 ± 6 M-1 min-1（试剂血红素基团在平行反应中的降解常数），ko = 51.0 ± 1.7 M-1 min-1（反应性双链体中靶标修饰常数）。由于修饰反应与试剂血红素基团在平行反应中的降解存在竞争，DNA的修饰并不完全。