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Protease‐resistant streptavidin for interaction proteomics
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 render...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7218406/ https://www.ncbi.nlm.nih.gov/pubmed/32400114 http://dx.doi.org/10.15252/msb.20199370 |
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author | Rafiee, Mahmoud‐Reza Sigismondo, Gianluca Kalxdorf, Mathias Förster, Laura Brügger, Britta Béthune, Julien Krijgsveld, Jeroen |
author_facet | Rafiee, Mahmoud‐Reza Sigismondo, Gianluca Kalxdorf, Mathias Förster, Laura Brügger, Britta Béthune, Julien Krijgsveld, Jeroen |
author_sort | Rafiee, Mahmoud‐Reza |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-7218406 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-72184062020-05-13 Protease‐resistant streptavidin for interaction proteomics Rafiee, Mahmoud‐Reza Sigismondo, Gianluca Kalxdorf, Mathias Förster, Laura Brügger, Britta Béthune, Julien Krijgsveld, Jeroen Mol Syst Biol Methods 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. John Wiley and Sons Inc. 2020-05-13 /pmc/articles/PMC7218406/ /pubmed/32400114 http://dx.doi.org/10.15252/msb.20199370 Text en © 2020 The Authors. Published under the terms of the CC BY 4.0 license This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methods Rafiee, Mahmoud‐Reza Sigismondo, Gianluca Kalxdorf, Mathias Förster, Laura Brügger, Britta Béthune, Julien Krijgsveld, Jeroen Protease‐resistant streptavidin for interaction proteomics |
title | 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_short | Protease‐resistant streptavidin for interaction proteomics |
title_sort | protease‐resistant streptavidin for interaction proteomics |
topic | Methods |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7218406/ https://www.ncbi.nlm.nih.gov/pubmed/32400114 http://dx.doi.org/10.15252/msb.20199370 |
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