Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies

Summary: We present a comprehensive and robust protocol to track the dynamics of all proteins in a complex in yeast cells. A single member of the protein assembly is tagged and conditionally expressed, minimizing the perturbations to the protein complex. Then, SILAC labeling and affinity purificatio...

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Main Authors:	Zhanna Hakhverdyan, Kelly R. Molloy, Roman I. Subbotin, Javier Fernandez-Martinez, Brian T. Chait, Michael P. Rout
Format:	Article
Language:	English
Published:	Elsevier 2021-09-01
Series:	STAR Protocols
Subjects:	Protein Biochemistry Proteomics
Online Access:	http://www.sciencedirect.com/science/article/pii/S2666166721005062

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spelling	doaj-8f14c84eb3e346c589f3e578b975f3cc2021-09-19T05:01:21ZengElsevierSTAR Protocols2666-16672021-09-0123100800Measuring in vivo protein turnover and exchange in yeast macromolecular assembliesZhanna Hakhverdyan0Kelly R. Molloy1Roman I. Subbotin2Javier Fernandez-Martinez3Brian T. Chait4Michael P. Rout5Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, NY 10065, USALaboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, NY 10065, USALaboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, NY 10065, USALaboratory of Cellular and Structural Biology, The Rockefeller University, New York, NY 10065, USA; Corresponding authorLaboratory of Cellular and Structural Biology, The Rockefeller University, New York, NY 10065, USA; Corresponding authorLaboratory of Cellular and Structural Biology, The Rockefeller University, New York, NY 10065, USA; Corresponding authorSummary: We present a comprehensive and robust protocol to track the dynamics of all proteins in a complex in yeast cells. A single member of the protein assembly is tagged and conditionally expressed, minimizing the perturbations to the protein complex. Then, SILAC labeling and affinity purification are used for the assessment of the whole protein complex dynamics. This method can determine and distinguish both subunit turnover and exchange specifically in an assembly to provide a comprehensive picture of assembly dynamics.For complete details on the use and execution of this protocol, please refer to Hakhverdyan et al. (2021).http://www.sciencedirect.com/science/article/pii/S2666166721005062Protein BiochemistryProteomics
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Zhanna Hakhverdyan Kelly R. Molloy Roman I. Subbotin Javier Fernandez-Martinez Brian T. Chait Michael P. Rout
spellingShingle	Zhanna Hakhverdyan Kelly R. Molloy Roman I. Subbotin Javier Fernandez-Martinez Brian T. Chait Michael P. Rout Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies STAR Protocols Protein Biochemistry Proteomics
author_facet	Zhanna Hakhverdyan Kelly R. Molloy Roman I. Subbotin Javier Fernandez-Martinez Brian T. Chait Michael P. Rout
author_sort	Zhanna Hakhverdyan
title	Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
title_short	Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
title_full	Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
title_fullStr	Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
title_full_unstemmed	Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
title_sort	measuring in vivo protein turnover and exchange in yeast macromolecular assemblies
publisher	Elsevier
series	STAR Protocols
issn	2666-1667
publishDate	2021-09-01
description	Summary: We present a comprehensive and robust protocol to track the dynamics of all proteins in a complex in yeast cells. A single member of the protein assembly is tagged and conditionally expressed, minimizing the perturbations to the protein complex. Then, SILAC labeling and affinity purification are used for the assessment of the whole protein complex dynamics. This method can determine and distinguish both subunit turnover and exchange specifically in an assembly to provide a comprehensive picture of assembly dynamics.For complete details on the use and execution of this protocol, please refer to Hakhverdyan et al. (2021).
topic	Protein Biochemistry Proteomics
url	http://www.sciencedirect.com/science/article/pii/S2666166721005062
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Measuring in vivo protein turnover and exchange in yeast macromolecular assemblies

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