Immunoaffinity purification of endogenous proteins from S. cerevisiae for post-translational modification and protein interaction analysis

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1170.pdf (2.25 MB)

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https://star-protocols.cell.com/protocols/1170

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https://doi.org/10.1016/j.xpro.2021.100945
 http://hdl.handle.net/11603/26706

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UMBC Biological Sciences Department
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Author/Creator ORCID

https://orcid.org/0000-0002-6514-7535
 https://orcid.org/0000-0003-3923-6726

Date

2021-11-19

Type of Work

journal articles
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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

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Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)

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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.