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

Summary: 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 an...

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Autores principales:	Deepika Jaiswal, Rashi Turniansky, Erin M. Green
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Model Organisms Signal Transduction Protein Biochemistry Proteomics Protein expression and purification Science (General) Q1-390
Acceso en línea:	https://doaj.org/article/a02a8b8b117e482487c9c01e5a50d630
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spelling	oai:doaj.org-article:a02a8b8b117e482487c9c01e5a50d6302021-11-14T04:35:36ZImmunoaffinity purification of endogenous proteins from S. cerevisiae for post-translational modification and protein interaction analysis2666-166710.1016/j.xpro.2021.100945https://doaj.org/article/a02a8b8b117e482487c9c01e5a50d6302021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2666166721006511https://doaj.org/toc/2666-1667Summary: 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.For complete details on the use and execution of this protocol, please refer to Jaiswal et al. (2020).Deepika JaiswalRashi TurnianskyErin M. GreenElsevierarticleModel OrganismsSignal TransductionProtein BiochemistryProteomicsProtein expression and purificationScience (General)Q1-390ENSTAR Protocols, Vol 2, Iss 4, Pp 100945- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Model Organisms Signal Transduction Protein Biochemistry Proteomics Protein expression and purification Science (General) Q1-390
spellingShingle	Model Organisms Signal Transduction Protein Biochemistry Proteomics Protein expression and purification Science (General) Q1-390 Deepika Jaiswal Rashi Turniansky Erin M. Green Immunoaffinity purification of endogenous proteins from S. cerevisiae for post-translational modification and protein interaction analysis
description	Summary: 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.For complete details on the use and execution of this protocol, please refer to Jaiswal et al. (2020).
format	article
author	Deepika Jaiswal Rashi Turniansky Erin M. Green
author_facet	Deepika Jaiswal Rashi Turniansky Erin M. Green
author_sort	Deepika Jaiswal
title	Immunoaffinity purification of endogenous proteins from S. cerevisiae for post-translational modification and protein interaction analysis
title_short	Immunoaffinity purification of endogenous proteins from S. cerevisiae for post-translational modification and protein interaction analysis
title_full	Immunoaffinity purification of endogenous proteins from S. cerevisiae for post-translational modification and protein interaction analysis
title_fullStr	Immunoaffinity purification of endogenous proteins from S. cerevisiae for post-translational modification and protein interaction analysis
title_full_unstemmed	Immunoaffinity purification of endogenous proteins from S. cerevisiae for post-translational modification and protein interaction analysis
title_sort	immunoaffinity purification of endogenous proteins from s. cerevisiae for post-translational modification and protein interaction analysis
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/a02a8b8b117e482487c9c01e5a50d630
work_keys_str_mv	AT deepikajaiswal immunoaffinitypurificationofendogenousproteinsfromscerevisiaeforposttranslationalmodificationandproteininteractionanalysis AT rashiturniansky immunoaffinitypurificationofendogenousproteinsfromscerevisiaeforposttranslationalmodificationandproteininteractionanalysis AT erinmgreen immunoaffinitypurificationofendogenousproteinsfromscerevisiaeforposttranslationalmodificationandproteininteractionanalysis
_version_	1718429914442498048

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

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