Protocol for Mapping Differential Protein-Protein Interaction Networks using Affinity Purification Mass Spectrometry

Autor: Kaushal, Prashant, Ummadi, Manisha R., Jang, Gwendolyn M., Delgado, Yennifer, Makanani, Sara K., Alba, Kareem, Winters, Decan M., Blanc, Sophie F., Xu, Jiewei, Polacco, Benjamin, Zhou, Yuan, Stevenson, Erica, Eckhardt, Manon, Zuliani-Alvarez, Lorena, Kaake, Robyn, Swaney, Danielle L., Krogan, Nevan, Bouhaddou, Mehdi
Zdroj: STAR Protocols; 20240101, Issue: Preprints
Abstrakt: 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 (APMS) 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.
Databáze: Supplemental Index