禽成髓细胞瘤病毒逆转录酶和大肠杆菌DNA聚合酶I的明显变构效应
Apparent allosterism by avian myeloblastosis virus reverse transcriptase and E. coli DNA polymerase I.
作者信息
Darling T L, Reid T W
出版信息
Nucleic Acids Res. 1979 Mar;6(3):1189-201. doi: 10.1093/nar/6.3.1189.
A recent report (1) presented evidence for allosterism in reverse transcription by Mason-Pfizer monkey virus reverse transcriptase and by E. coli DNA polymerase I. Our experiments also demonstrate these apparent cooperative effects when synthesis is catalyzed by either avian myeloblastosis virus DNA polymerase, feline sarcoma virus DNA polymerase, or E. coli DNA polymerase I (large fragment). We show that the apparent cooperativity depends on the use of oligo(dT)12-18 as primer. However, if the polymerase reaction products are isolated chromatographically, then the polymerases obey classical Michaelis-Menten kinetics with respect to substrate and enzyme concentrations. These results suggest that the cooperative effects are an acid precipitation artifact. The results are also consistent with the enzyme operating by a distributive mechanism with the oligo(dT)12-18 primer.
最近的一份报告（1）提供了证据，证明梅森- Pfizer猴病毒逆转录酶和大肠杆菌DNA聚合酶I在逆转录过程中存在变构现象。我们的实验也证明，当由禽成髓细胞瘤病毒DNA聚合酶、猫肉瘤病毒DNA聚合酶或大肠杆菌DNA聚合酶I（大片段）催化合成时，也会出现这些明显的协同效应。我们表明，这种明显的协同性取决于使用寡聚（dT）12 - 18作为引物。然而，如果通过色谱法分离聚合酶反应产物，那么这些聚合酶在底物和酶浓度方面遵循经典的米氏动力学。这些结果表明，协同效应是酸沉淀假象。这些结果也与该酶通过寡聚（dT）12 - 18引物以分布机制起作用相一致。
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