Tam Annie, Raines Ronald T
Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Methods Enzymol. 2009;462:25-44. doi: 10.1016/S0076-6879(09)62002-4.
The engineering of proteins can illuminate their biological function and improve their performance in a variety of applications. Within the past decade, methods have been developed that facilitate the ability of chemists to manipulate proteins in a controlled manner. Here, we present the traceless Staudinger ligation as a strategy for the convergent chemical synthesis of proteins. This reaction unites a phosphinothioester and an azide to form an amide bond with no residual atoms. An important feature of this reaction is its ability to ligate peptides at noncysteine residues, thereby overcoming a limitation of alternative strategies. Attributes of the traceless Staudinger ligation are discussed, and an overall comparison of known reagents for effecting the reaction is presented. General methods are elaborated for the synthesis of the most efficacious phosphinothiol for mediating the traceless Staudinger ligation, as well as for the preparation of phosphinothioester and azide fragments and the ligation of peptides immobilized on a solid support. Together, this information facilitates the use of this emerging method to engineer proteins.
蛋白质工程能够阐明其生物学功能，并在各种应用中提升其性能。在过去十年间，已开发出一些方法，有助于化学家以可控方式操纵蛋白质。在此，我们介绍无痕施陶丁格连接反应，作为一种用于蛋白质收敛化学合成的策略。该反应将磷硫酯和叠氮化物结合形成酰胺键，且无残留原子。此反应的一个重要特性是其能够在非半胱氨酸残基处连接肽段，从而克服了其他策略的一个局限性。文中讨论了无痕施陶丁格连接反应的特性，并对用于实现该反应的已知试剂进行了全面比较。详细阐述了合成用于介导无痕施陶丁格连接反应的最有效磷硫醇的通用方法，以及磷硫酯和叠氮化物片段的制备方法，以及固定在固体支持物上的肽段的连接方法。这些信息共同促进了这种新兴方法在蛋白质工程中的应用。
