Protocol for establishing a protein interactome based on close physical proximity to a target protein within live budding yeast

Janhavi A. Kolhe, Neethu L. Babu, Brian C. Freeman

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish (US)
Article number102663
JournalSTAR Protocols
Volume4
Issue number4
DOIs
StatePublished - Dec 15 2023

Keywords

  • Model Organisms
  • Molecular Biology
  • Protein Biochemistry

ASJC Scopus subject areas

  • General Immunology and Microbiology
  • General Biochemistry, Genetics and Molecular Biology
  • General Neuroscience

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