Fast-scanning atomic force microscopy reveals the molecular mechanism of DNA cleavage by ApaI endonuclease.

M Yokokawa, S H Yoshimura, Y Naito, T Ando, A Yagi, N Sakai, K Takeyasu
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引用次数: 5

Abstract

Newly developed fast-scanning atomic force microscopy (AFM) allows the dissection of molecular events such as DNA-enzyme reactions at the single-molecule level. With this novel technology, a model is proposed of the DNA cleavage reaction by a type IIP restriction endonuclease ApaI. Detailed analyses revealed that ApaI bound to DNA as a dimer and slid along DNA in a one-dimensional diffusion manner. When it encountered a specific DNA sequence, the enzyme halted for a moment to digest the DNA. Immediately after digestion, the ApaI dimer separated into two monomers, each of which remained on the DNA end and then dissociated from the DNA end. Thus, fast-scanning AFM is a powerful tool to aid the understanding of protein structures and dynamics in biological reactions at the single-molecule level in sub-seconds.

快速扫描原子力显微镜揭示了ApaI内切酶裂解DNA的分子机制。
新开发的快速扫描原子力显微镜(AFM)允许在单分子水平上解剖分子事件,如dna -酶反应。利用这种新技术,提出了IIP型限制性内切酶ApaI的DNA切割反应模型。详细分析表明,ApaI以二聚体形式与DNA结合,并以一维扩散方式沿着DNA滑动。当它遇到特定的DNA序列时,酶会暂停片刻来消化DNA。消化后,ApaI二聚体立即分离成两个单体,每个单体都留在DNA端,然后从DNA端解离。因此,快速扫描AFM是一种强大的工具,可以帮助理解单分子水平生物反应中的蛋白质结构和动力学。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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