Expressed protein ligation (EPL), using non–self-cleaving inteins, allows for the site-specific addition of customized chemical moieties to the termini of proteins. In this way, protein activity can be preserved while functionalizing the target protein with a wide range of chemical handles. Here, we describe methods for EPL-based modification of proteins produced by yeast, employing an engineered, non–self-cleaving intein known as 202-08. Methods for EPL modification of both yeast surface displayed and secreted proteins with bioorthogonal chemical groups are described. These methods allow for the site-specific modification of intein-fused proteins produced in yeast.
CITATION STYLE
Umlauf, B. J., & Shusta, E. V. (2020). Site-Directed Modification of Yeast-Produced Proteins Using Expressed Protein Ligation. In Methods in Molecular Biology (Vol. 2133, pp. 221–233). Humana Press Inc. https://doi.org/10.1007/978-1-0716-0434-2_11
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