University of Pennsylvania, Philadelphia, PA, USA.
Methods Mol Biol. 2023;2620:177-207. doi: 10.1007/978-1-0716-2942-0_22.
Solid-phase peptide synthesis and protein semi-synthesis are powerful methods for site-specific modification of peptides and proteins. We describe protocols using these techniques for the syntheses of peptides and proteins bearing glutamate arginylation (E) at specific sites. These methods overcome challenges posed by enzymatic arginylation methods and allow for a comprehensive study of the effects of E on protein folding and interactions. Potential applications include biophysical analyses, cell-based microscopic studies, and profiling of E levels and interactomes in human tissue samples.
固相肽合成和蛋白质半合成是对肽和蛋白质进行定点修饰的有力方法。我们描述了使用这些技术在特定位点合成带有谷氨酸精氨酸化（E）的肽和蛋白质的方案。这些方法克服了酶促精氨酸化方法带来的挑战，并且可以全面研究 E 对蛋白质折叠和相互作用的影响。潜在的应用包括生物物理分析、基于细胞的显微镜研究，以及在人体组织样本中分析 E 水平和相互作用组。
