School of Science, Western Sydney University, Penrith, NSW, Australia.
School of Life and Environmental Sciences, University of Sydney, Sydney, NSW, Australia.
Methods Mol Biol. 2021;2281:229-240. doi: 10.1007/978-1-0716-1290-3_14.
Single-stranded DNA-binding proteins (SSBs) are essential to all living organisms as protectors and guardians of the genome. Apart from the well-characterized RPA, humans have also evolved two further SSBs, termed hSSB1 and hSSB2. Over the last few years, we have used NMR spectroscopy to determine the molecular and structural details of both hSSBs and their interactions with DNA and RNA. Here we provide a detailed overview of how to express and purify recombinant versions of these important human proteins for the purpose of detailed structural analysis by high-resolution solution-state NMR.
单链 DNA 结合蛋白（SSBs）作为基因组的保护者和守护者，对所有生物都是必不可少的。除了众所周知的 RPA 之外，人类还进化出了另外两种 SSB，分别称为 hSSB1 和 hSSB2。在过去的几年中，我们使用 NMR 光谱学来确定这两种 hSSB 及其与 DNA 和 RNA 的相互作用的分子和结构细节。在这里，我们提供了一个详细的概述，介绍如何表达和纯化这些重要的人类蛋白质的重组版本，以便通过高分辨率溶液 NMR 进行详细的结构分析。
