Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX, USA.
Department of Chemistry, Texas A&M University, College Station, TX, USA.
Methods Mol Biol. 2020;2133:75-117. doi: 10.1007/978-1-0716-0434-2_5.
Expressed protein ligation allows for the attachment of a chemically labeled peptide to the N- or C-terminus of a recombinant protein. In this book chapter, the practical considerations involved in using this protein engineering technology are described. In particular, approaches used to design optimal ligation sites are discussed. In addition, several methods used to generate the reactive fragments required for EPL are highlighted in practical details. Finally, strategies that one can implement to achieve efficient ligation reactions are presented.
蛋白定点连接允许将化学标记的肽连接到重组蛋白的 N 端或 C 端。在本章中,描述了使用这项蛋白质工程技术所涉及的实际考虑因素。特别是,讨论了用于设计最佳连接位点的方法。此外,还详细介绍了几种用于生成 EPL 所需的反应性片段的方法。最后,提出了实现高效连接反应的策略。