Protocol for establishing a protein interactome based on close physical proximity to a target protein within live budding yeast
Summary: Here, we present a protocol for establishing a protein interactome based on close physical proximity to a target protein within live yeast cells. We describe steps for capturing both transient and stable binders by integrating a non-natural amino acid. We detail procedures for employing a s...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723006305 |
| _version_ | 1797648354339979264 |
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| author | Janhavi A. Kolhe Neethu L. Babu Brian C. Freeman |
| author_facet | Janhavi A. Kolhe Neethu L. Babu Brian C. Freeman |
| author_sort | Janhavi A. Kolhe |
| collection | DOAJ |
| description | Summary: Here, we present a protocol for establishing a protein interactome based on close physical proximity to a target protein within live yeast cells. We describe steps for capturing both transient and stable binders by integrating a non-natural amino acid. We detail procedures for employing a site-directed method for labeling the surface that mediates protein associations and uncovers the binding sites on the interactors. Combined with mass spectrometry, our approach proves valuable in discovering binding partners and constructing a comprehensive protein-interaction network. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-11T15:31:02Z |
| format | Article |
| id | doaj.art-419e2e9a3dae4ad5ad463e4c68f18ce2 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-11T15:31:02Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-419e2e9a3dae4ad5ad463e4c68f18ce22023-10-27T04:24:41ZengElsevierSTAR Protocols2666-16672023-12-0144102663Protocol for establishing a protein interactome based on close physical proximity to a target protein within live budding yeastJanhavi A. Kolhe0Neethu L. Babu1Brian C. Freeman2Department of Cell and Developmental Biology, School of Molecular and Cellular Biology, University of Illinois-Urbana-Champaign, Urbana, IL, USADepartment of Cell and Developmental Biology, School of Molecular and Cellular Biology, University of Illinois-Urbana-Champaign, Urbana, IL, USADepartment of Cell and Developmental Biology, School of Molecular and Cellular Biology, University of Illinois-Urbana-Champaign, Urbana, IL, USA; Corresponding authorSummary: Here, we present a protocol for establishing a protein interactome based on close physical proximity to a target protein within live yeast cells. We describe steps for capturing both transient and stable binders by integrating a non-natural amino acid. We detail procedures for employing a site-directed method for labeling the surface that mediates protein associations and uncovers the binding sites on the interactors. Combined with mass spectrometry, our approach proves valuable in discovering binding partners and constructing a comprehensive protein-interaction network. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723006305Model OrganismsMolecular BiologyProtein Biochemistry |
| spellingShingle | Janhavi A. Kolhe Neethu L. Babu Brian C. Freeman Protocol for establishing a protein interactome based on close physical proximity to a target protein within live budding yeast STAR Protocols Model Organisms Molecular Biology Protein Biochemistry |
| title | Protocol for establishing a protein interactome based on close physical proximity to a target protein within live budding yeast |
| title_full | Protocol for establishing a protein interactome based on close physical proximity to a target protein within live budding yeast |
| title_fullStr | Protocol for establishing a protein interactome based on close physical proximity to a target protein within live budding yeast |
| title_full_unstemmed | Protocol for establishing a protein interactome based on close physical proximity to a target protein within live budding yeast |
| title_short | Protocol for establishing a protein interactome based on close physical proximity to a target protein within live budding yeast |
| title_sort | protocol for establishing a protein interactome based on close physical proximity to a target protein within live budding yeast |
| topic | Model Organisms Molecular Biology Protein Biochemistry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723006305 |
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