A proteomics protocol to identify stimulation-induced binding partners dependent on a specific gene in mammalian cells

Summary: Some protein-protein interactions are induced by different kinds of stimulation and are dependent on specific genes. To identify these interaction partners, we present a protocol which utilizes affinity purification of Flag-tagged protein complexes followed by mass-spectrometry-based proteo...

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Autores principales:	Junji Zhu, Wuhan Xiao
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Chromatin immunoprecipitation (ChIP) CRISPR Mass Spectrometry Protein Biochemistry Protein expression and purification Proteomics Science (General) Q1-390
Acceso en línea:	https://doaj.org/article/5b067d16d3214e3b9d2f7d55f45809db
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spelling	oai:doaj.org-article:5b067d16d3214e3b9d2f7d55f45809db2021-11-14T04:35:39ZA proteomics protocol to identify stimulation-induced binding partners dependent on a specific gene in mammalian cells2666-166710.1016/j.xpro.2021.100962https://doaj.org/article/5b067d16d3214e3b9d2f7d55f45809db2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2666166721006687https://doaj.org/toc/2666-1667Summary: Some protein-protein interactions are induced by different kinds of stimulation and are dependent on specific genes. To identify these interaction partners, we present a protocol which utilizes affinity purification of Flag-tagged protein complexes followed by mass-spectrometry-based proteomics to compare stimulation-induced interactomes between wild-type and CRISPR-Cas9-mediated knockout cells. The candidates of interest are identified using bioinformatic analyses and verified by biochemical approaches. This protocol is highly versatile and applies to a variety of cells and different types of stimulation.For complete details on the use and execution of this protocol, please refer to (Zhu et al., 2021).Junji ZhuWuhan XiaoElsevierarticleChromatin immunoprecipitation (ChIP)CRISPRMass SpectrometryProtein BiochemistryProtein expression and purificationProteomicsScience (General)Q1-390ENSTAR Protocols, Vol 2, Iss 4, Pp 100962- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Chromatin immunoprecipitation (ChIP) CRISPR Mass Spectrometry Protein Biochemistry Protein expression and purification Proteomics Science (General) Q1-390
spellingShingle	Chromatin immunoprecipitation (ChIP) CRISPR Mass Spectrometry Protein Biochemistry Protein expression and purification Proteomics Science (General) Q1-390 Junji Zhu Wuhan Xiao A proteomics protocol to identify stimulation-induced binding partners dependent on a specific gene in mammalian cells
description	Summary: Some protein-protein interactions are induced by different kinds of stimulation and are dependent on specific genes. To identify these interaction partners, we present a protocol which utilizes affinity purification of Flag-tagged protein complexes followed by mass-spectrometry-based proteomics to compare stimulation-induced interactomes between wild-type and CRISPR-Cas9-mediated knockout cells. The candidates of interest are identified using bioinformatic analyses and verified by biochemical approaches. This protocol is highly versatile and applies to a variety of cells and different types of stimulation.For complete details on the use and execution of this protocol, please refer to (Zhu et al., 2021).
format	article
author	Junji Zhu Wuhan Xiao
author_facet	Junji Zhu Wuhan Xiao
author_sort	Junji Zhu
title	A proteomics protocol to identify stimulation-induced binding partners dependent on a specific gene in mammalian cells
title_short	A proteomics protocol to identify stimulation-induced binding partners dependent on a specific gene in mammalian cells
title_full	A proteomics protocol to identify stimulation-induced binding partners dependent on a specific gene in mammalian cells
title_fullStr	A proteomics protocol to identify stimulation-induced binding partners dependent on a specific gene in mammalian cells
title_full_unstemmed	A proteomics protocol to identify stimulation-induced binding partners dependent on a specific gene in mammalian cells
title_sort	proteomics protocol to identify stimulation-induced binding partners dependent on a specific gene in mammalian cells
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/5b067d16d3214e3b9d2f7d55f45809db
work_keys_str_mv	AT junjizhu aproteomicsprotocoltoidentifystimulationinducedbindingpartnersdependentonaspecificgeneinmammaliancells AT wuhanxiao aproteomicsprotocoltoidentifystimulationinducedbindingpartnersdependentonaspecificgeneinmammaliancells AT junjizhu proteomicsprotocoltoidentifystimulationinducedbindingpartnersdependentonaspecificgeneinmammaliancells AT wuhanxiao proteomicsprotocoltoidentifystimulationinducedbindingpartnersdependentonaspecificgeneinmammaliancells
_version_	1718429898082615296

A proteomics protocol to identify stimulation-induced binding partners dependent on a specific gene in mammalian cells

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