The inhibition of deoxyribonucleotidyl transferase, DNAase and RNAase by sodium poly ethenesulfonic acid. Effect of the molecular weight of the inhibitor

Michael K. Bach
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引用次数: 21

Abstract

  • 1.

    1. The nature of the inhibition of DNA nucleotidyltransferase (EC 2.7.7.7) from KB cells (DNA polymerase) by poly ethenesulfonic acid (PES) and other polyanionic materials were examined. The inhibition was found to be competitive with the DNA used to prime the enzyme. The K1 was found to be 1.72 ± 0.36 μg PES/ml for the 12 900 mol. wt. material in the range of zero to 10 μg inhibitor/ml. The inhibitory action of PES was strongly dependent on the molecular weight of the sample used in the range of 5000 to 13 000 and further increases in molecular weight did not seem to affect efficacy. Other polyanionic polymers, such as heparin, carboxymethyl cellulose, alginic acid, chitosan sulfate, RNA and chondroitin sulfate also inhibited this enzyme to varying degrees.

  • 2.

    2. Spermine, which by itself inhibits the DNA polymerase, reversed the inhibition caused by PES. This is explained by the competition between DNA and PES for salt formation (and precipitation) with spermine.

  • 3.

    3. The inhibition of pancreatic RNAase (EC 2.7.7.16) by PES exhibited a similar dependence on the molecular weight of the polymer as did the inhibition of DNA polymerase.

  • 4.

    4. Addition of PES to DNAase I (EC 3.1.4.5) assays resulted in a bimodal, concentration-dependent curve. At intermediate concentrations the compound stimulated DNA hydrolysis, while at higher concentrations it inhibited.

聚乙烯磺酸钠对脱氧核糖核酸转移酶、脱氧核糖核酸酶和核糖核酸酶的抑制作用。抑制剂分子量的影响
1.1. 研究了聚乙烯磺酸(PES)和其他聚阴离子材料对KB细胞DNA聚合酶(DNA nucleotidyltransferase, EC 2.7.7.7)的抑制作用。抑制作用被发现与用于引物酶的DNA竞争。在0 ~ 10 μg抑制剂/ml范围内,12 900 mol. wt材料的K1为1.72±0.36 μg PES/ml。PES的抑制作用强烈依赖于所使用样品的分子量,在5000至13000范围内,分子量的进一步增加似乎不影响效果。其他聚阴离子聚合物,如肝素、羧甲基纤维素、海藻酸、硫酸壳聚糖、RNA和硫酸软骨素也不同程度地抑制该酶。精胺本身可以抑制DNA聚合酶,逆转了PES引起的抑制作用。这可以解释为DNA和PES在与精胺形成盐(和沉淀)方面的竞争。PES对胰腺RNAase (EC 2.7.7.16)的抑制作用与对DNA聚合酶的抑制作用同样依赖于聚合物的分子量。将PES添加到DNAase I (EC 3.1.4.5)检测中,产生双峰浓度依赖性曲线。在中等浓度下,该化合物刺激DNA水解,而在较高浓度下则抑制DNA水解。
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