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Reverse transcriptase: a monitor for perturbation effects of spin labels covalently bound to nucleic acids.

机译:逆转录酶:用于监控与核酸共价结合的自旋标记的干扰作用的监控器。

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摘要

The sensitive biological assay for reverse transcriptase was used to monitor potential perturbation effects of spin labels covalently bound to various nucleic acids or nucleic acid analogs to the extent of about one label per 100 residues. The inhibitory properties of the spin labeled and unlabeled biopolymers were compared for evaluating possible interference of the reporter group in protein-nucleic acid interaction studies. The amount of inhibitor required for 50% inhibition (ED50) was determined for the competitive inhibitors (U)n, l(U)n, (RUGT,U)n, (Um)n, (A)n, (Am)n, and l(A)n as well as for thenon- or uncompetitive inhibitors (dUfl)n, l(dUfl)n, (dUz)n, and l(dUz)n. The most pronounced inhibition was observed with spin labeled and unlabeled (dUfl)n. The results indicate that the ED50 and the kinetic patterns of inhibition are similar for the spin labeled and unlabeled inhibitors studied. Thus, the presence of a limited number of spin labels in a nucleic acid matrix has little effect, if any, on reverse transcriptase-nucleic acid complexes and most likely on other protein-nucleic acid complexes.
机译:用于逆转录酶的灵敏生物测定法用于监测共价结合至各种核酸或核酸类似物的自旋标记的潜在扰动效应,其程度约为每100个残基一个标记。比较了自旋标记的和未标记的生物聚合物的抑制特性,以评估报告基团在蛋白质-核酸相互作用研究中的可能干扰。确定竞争性抑制剂(U)n,l(U)n,(RUGT,U)n,(Um)n,(A)n,(Am)n的50%抑制所需的抑制剂量(ED50) ,l(A)n以及非竞争性或非竞争性抑制剂(dUfl)n,l(dUfl)n,(dUz)n和l(dUz)n。用旋转标记的和未标记的(dUf1)n观察到最明显的抑制作用。结果表明,所研究的旋转标记和未标记抑制剂的ED50和抑制动力学模式相似。因此,核酸基质中有限数量的自旋标记的存在(如果有的话)对逆转录酶-核酸复合物几乎没有影响,而对其他蛋白质-核酸复合物的影响则很小。

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