Department of Chemistry, East Carolina University, Greenville, NC, United States.
Department of Chemistry, East Carolina University, Greenville, NC, United States.
Methods Enzymol. 2021;647:83-106. doi: 10.1016/bs.mie.2020.10.001. Epub 2020 Nov 9.
The impacts of linkers on dynamics, expression, and activity of biomacromolecules are often overlooked. This may be due, in part, to the lack of facile methods for incorporation and analysis of linkers that vary iteratively in both length and sequence composition. The protaTETHER method addresses this gap by enabling the incorporation of focused linker libraries at potentially any region in a protein sequence. In this chapter, we describe the generation and incorporation of linkers in a PKAc-GFP fusion protein and provide methods for the application and evaluation of the protaTETHER process.
链接物对生物大分子的动力学、表达和活性的影响经常被忽视。这可能部分归因于缺乏方便的方法来整合和分析在长度和序列组成上都不断变化的链接物。protaTETHER 方法通过在蛋白质序列的潜在任何区域中引入聚焦的链接物文库来解决这一差距。在本章中，我们描述了在 PKAc-GFP 融合蛋白中生成和引入链接物的方法，并提供了应用和评估 protaTETHER 过程的方法。
