Immunoaffinity purification of endogenous proteins from S. cerevisiae for post-translational modification and protein interaction analysis
Files
Links to Files
Author/Creator
Author/Creator ORCID
Date
Type of Work
Department
Program
Citation of Original Publication
Jaiswal, Deepika, Rashi Turniansky and Erin M. Green. “Immunoaffinity purification of endogenous proteins from S. cerevisiae for post-translational modification and protein interaction analysis.” STAR Protocols 2, 100945 (December 17, 2021). https://doi.org/10.1016/j.xpro.2021.100945
Rights
Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
This item is likely protected under Title 17 of the U.S. Copyright Law. Unless on a Creative Commons license, for uses protected by Copyright Law, contact the copyright holder or the author.
This item is likely protected under Title 17 of the U.S. Copyright Law. Unless on a Creative Commons license, for uses protected by Copyright Law, contact the copyright holder or the author.
Subjects
Abstract
Protein regulation by post-translational modifications and protein-protein interactions is critical to controlling molecular pathways. Here, we describe an immunoaffinity purification approach in Saccharomyces cerevisiae. The protocol uses an endogenously-expressed epitope-tagged protein and can be applied to the identification of post-translational modifications or protein binding partners. The lysine methyltransferase Set5 is used as an example here to purify phosphorylated Set5 and identify phosphosites; however, this approach can be applied to a diverse set of proteins in yeast.