Protocol for mapping differential protein-protein interaction networks using affinity purification-mass spectrometry.

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  • Additional Information
    • Source:
      Publisher: Cell Press Country of Publication: United States NLM ID: 101769501 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2666-1667 (Electronic) Linking ISSN: 26661667 NLM ISO Abbreviation: STAR Protoc Subsets: MEDLINE
    • Publication Information:
      Original Publication: [Cambridge, MA] : Cell Press, [2020]-
    • Subject Terms:
      Protein Interaction Mapping*/methods ; Mass Spectrometry*/methods ; Chromatography, Affinity*/methods ; Protein Interaction Maps*/physiology ; Proteomics*/methods; Humans ; Proteins/metabolism ; HEK293 Cells
    • Abstract:
      Proteins congregate into complexes to perform diverse cellular functions. Protein complexes are remodeled by protein-coding mutations or cellular signaling changes, driving phenotypic outcomes in health and disease. We present an affinity purification-mass spectrometry (AP-MS) proteomics protocol to express affinity-tagged "bait" proteins in mammalian cells, identify and quantify purified protein interactors, and visualize differential protein-protein interaction networks between pairwise conditions. Our protocol possesses general applicability to various cell types and biological areas. For complete details on the use and execution of this protocol, please refer to Bouhaddou et al. 1 .
      Competing Interests: Declaration of interests The Krogan Laboratory has received research support from Vir Biotechnology, F. Hoffmann-La Roche, and Rezo Therapeutics. N.J.K. has a financially compensated consulting agreement with Maze Therapeutics. N.J.K. is the President and is on the Board of Directors of Rezo Therapeutics, and he is a shareholder in Tenaya Therapeutics, Maze Therapeutics, Rezo Therapeutics, and Interline Therapeutics. M.B. is a scientific advisor for GEn1E LifeSciences.
      (Copyright © 2024. Published by Elsevier Inc.)
    • Comments:
      Update of: ArXiv. 2024 May 15:arXiv:2405.09699v1.. (PMID: 38800652)
    • Contributed Indexing:
      Keywords: Mass Spectrometry; Molecular Biology; Protein Biochemistry; Protein expression and purification; Proteomics; Systems biology
    • Accession Number:
      0 (Proteins)
    • Publication Date:
      Date Created: 20241103 Date Completed: 20241224 Latest Revision: 20241224
    • Publication Date:
      20241224
    • Accession Number:
      PMC11567037
    • Accession Number:
      10.1016/j.xpro.2024.103286
    • Accession Number:
      39488835