Department of Chemistry, College of Staten Island, CUNY, Staten Island, NY 10314, USA.
J Pept Sci. 2010 May;16(5):213-8. doi: 10.1002/psc.1222.
Biosynthesis of peptides in heterologous systems is often a prerequisite to biophysical analyses. Large amounts of peptides, in particular portions of membrane proteins, are needed to optimize conditions for secondary and tertiary structure analysis. Chemical synthesis of these peptides is limited by their high hydrophobicity and also due to the need to incorporate isotopic labels for high resolution NMR analysis. In this protocol, we describe a method for the heterologous expression and purification of unlabeled and isotopically labeled peptide fragments of Ste2p, an integral membrane G protein-coupled receptor.
在异源系统中进行肽的生物合成通常是生物物理分析的前提。需要大量的肽，特别是膜蛋白的部分，以优化二级和三级结构分析的条件。这些肽的化学合成受到其高度疏水性的限制，并且还需要掺入同位素标记物以进行高分辨率 NMR 分析。在本方案中，我们描述了一种用于异源表达和纯化 Ste2p 的未标记和同位素标记肽片段的方法，Ste2p 是一种完整的膜 G 蛋白偶联受体。
