414 Wartik Laboratories, Department of Chemistry, The Pennsylvania State University, University Park, PA 16802, USA.
Methods. 2010 Jul;51(3):277-88. doi: 10.1016/j.ymeth.2010.02.011. Epub 2010 Feb 17.
Helicases are an important class of enzymes involved in DNA and RNA metabolism that couple the energy of ATP hydrolysis to unwind duplex DNA and RNA structures. Understanding the mechanism of helicase action is vital due to their involvement in various biological processes such as DNA replication, repair and recombination. Furthermore, the duplex DNA unwinding property of this class of enzymes is closely related to their single-stranded DNA translocation. Hence the study of its translocation properties is essential to understanding helicase activity. Here we review the methods that are employed to analyze the DNA translocation and unwinding activities of the bacteriophage T4 UvsW and Dda helicases. These methods have been successfully employed to study the functions of helicases from large superfamilies.
解旋酶是一类在 DNA 和 RNA 代谢中起重要作用的酶,它将 ATP 水解的能量耦合到双链 DNA 和 RNA 结构的解旋中。由于它们参与各种生物过程,如 DNA 复制、修复和重组,因此了解解旋酶作用的机制至关重要。此外,这类酶的双链 DNA 解旋特性与其单链 DNA 易位密切相关。因此,研究其易位特性对于理解解旋酶的活性至关重要。在这里,我们回顾了用于分析噬菌体 T4 UvsW 和 Dda 解旋酶的 DNA 易位和解旋活性的方法。这些方法已成功地用于研究来自大型超家族的解旋酶的功能。