Protease‐resistant streptavidin for interaction proteomics

Protease‐resistant streptavidin for interaction proteomics

Abstract Streptavidin‐mediated enrichment is a powerful strategy to identify biotinylated biomolecules and their interaction partners; however, intense streptavidin‐derived peptides impede protein identification by mass spectrometry. Here, we present an approach to chemically modify streptavidin, th...

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Main Authors: Mahmoud‐Reza Rafiee, Gianluca Sigismondo, Mathias Kalxdorf, Laura Förster, Britta Brügger, Julien Béthune, Jeroen Krijgsveld
Format: Article
Language:English
Published: Wiley 2020-05-01
Series:Molecular Systems Biology
Subjects:
chemical derivatization
ChIP‐SICAP
contamination
proteomics
streptavidin
Online Access:https://doi.org/10.15252/msb.20199370
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spelling doaj-38d85c958ac1461aaa30bac0a429cf0c2021-08-02T12:03:07ZengWileyMolecular Systems Biology1744-42922020-05-01165n/an/a10.15252/msb.20199370Protease‐resistant streptavidin for interaction proteomicsMahmoud‐Reza Rafiee0Gianluca Sigismondo1Mathias Kalxdorf2Laura Förster3Britta Brügger4Julien Béthune5Jeroen Krijgsveld6Division of Proteomics of Stem Cells and Cancer German Cancer Research Center (DKFZ) Heidelberg GermanyDivision of Proteomics of Stem Cells and Cancer German Cancer Research Center (DKFZ) Heidelberg GermanyDivision of Proteomics of Stem Cells and Cancer German Cancer Research Center (DKFZ) Heidelberg GermanyHeidelberg University Biochemistry Center (BZH) Heidelberg GermanyHeidelberg University Biochemistry Center (BZH) Heidelberg GermanyHeidelberg University Biochemistry Center (BZH) Heidelberg GermanyDivision of Proteomics of Stem Cells and Cancer German Cancer Research Center (DKFZ) Heidelberg GermanyAbstract Streptavidin‐mediated enrichment is a powerful strategy to identify biotinylated biomolecules and their interaction partners; however, intense streptavidin‐derived peptides impede protein identification by mass spectrometry. Here, we present an approach to chemically modify streptavidin, thus rendering it resistant to proteolysis by trypsin and LysC. This modification results in over 100‐fold reduction of streptavidin contamination and in better coverage of proteins interacting with various biotinylated bait molecules (DNA, protein, and lipid) in an overall simplified workflow.https://doi.org/10.15252/msb.20199370chemical derivatizationChIP‐SICAPcontaminationproteomicsstreptavidin
collection DOAJ
language English
format Article
sources DOAJ
author Mahmoud‐Reza Rafiee
Gianluca Sigismondo
Mathias Kalxdorf
Laura Förster
Britta Brügger
Julien Béthune
Jeroen Krijgsveld
spellingShingle Mahmoud‐Reza Rafiee
Gianluca Sigismondo
Mathias Kalxdorf
Laura Förster
Britta Brügger
Julien Béthune
Jeroen Krijgsveld
Protease‐resistant streptavidin for interaction proteomics
Molecular Systems Biology
chemical derivatization
ChIP‐SICAP
contamination
proteomics
streptavidin
author_facet Mahmoud‐Reza Rafiee
Gianluca Sigismondo
Mathias Kalxdorf
Laura Förster
Britta Brügger
Julien Béthune
Jeroen Krijgsveld
author_sort Mahmoud‐Reza Rafiee
title Protease‐resistant streptavidin for interaction proteomics
title_short Protease‐resistant streptavidin for interaction proteomics
title_full Protease‐resistant streptavidin for interaction proteomics
title_fullStr Protease‐resistant streptavidin for interaction proteomics
title_full_unstemmed Protease‐resistant streptavidin for interaction proteomics
title_sort protease‐resistant streptavidin for interaction proteomics
publisher Wiley
series Molecular Systems Biology
issn 1744-4292
publishDate 2020-05-01
description Abstract Streptavidin‐mediated enrichment is a powerful strategy to identify biotinylated biomolecules and their interaction partners; however, intense streptavidin‐derived peptides impede protein identification by mass spectrometry. Here, we present an approach to chemically modify streptavidin, thus rendering it resistant to proteolysis by trypsin and LysC. This modification results in over 100‐fold reduction of streptavidin contamination and in better coverage of proteins interacting with various biotinylated bait molecules (DNA, protein, and lipid) in an overall simplified workflow.
topic chemical derivatization
ChIP‐SICAP
contamination
proteomics
streptavidin
url https://doi.org/10.15252/msb.20199370
work_keys_str_mv AT mahmoudrezarafiee proteaseresistantstreptavidinforinteractionproteomics
AT gianlucasigismondo proteaseresistantstreptavidinforinteractionproteomics
AT mathiaskalxdorf proteaseresistantstreptavidinforinteractionproteomics
AT lauraforster proteaseresistantstreptavidinforinteractionproteomics
AT brittabrugger proteaseresistantstreptavidinforinteractionproteomics
AT julienbethune proteaseresistantstreptavidinforinteractionproteomics
AT jeroenkrijgsveld proteaseresistantstreptavidinforinteractionproteomics
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