title: Sample Preparation for Mass Spectrometry Analysis of Protein-Protein Interactions in Cancer Cell Lines and Tissues.
abstract: A precisely controlled network of protein-protein interactions constitutes the basis for functional signaling pathways. This equilibrium is more often than not disrupted in cancer cells, by the aberrant expression or activation of oncogenic proteins. Therefore, the analysis of protein interaction networks in cancer cells has become crucial to expand our comprehension of the molecular underpinnings of tumor formation and progression. This protocol describes a sample preparation method for the analysis of signaling complexes by mass spectrometry (MS), following the affinity purification of a protein of interest from a cancer cell line or a solid tumor. In particular, we provide a spin tip-based protease digestion procedure that offers a more rapid and controlled alternative to other gel-based and gel-free methods. This sample preparation protocol represents a useful strategy to identify protein interactions and to gain insight into the molecular mechanisms that contribute to a given cancer phenotype.
authors: Division of Oncology, Centre de recherche du CHU de Québec, Québec, QC, Canada.|Cancer Research Centre, Université Laval, Québec, QC, Canada.
authors: Alice
authors: A
authors: Beigbeder
authors: Division of Oncology, Centre de recherche du CHU de Québec, Québec, QC, Canada.|Cancer Research Centre, Université Laval, Québec, QC, Canada.
authors: Lauriane
authors: L
authors: Vélot
authors: Department of Chemistry, York University, Toronto, ON, Canada.
authors: D Andrew
authors: DA
authors: James
authors: Division of Oncology, Centre de recherche du CHU de Québec, Québec, QC, Canada. nick.bisson@crchudequebec.ulaval.ca.|Cancer Research Centre, Université Laval, Québec, QC, Canada. nick.bisson@crchudequebec.ulaval.ca.|Quebec Network for Research on Protein Function, Engineering, and Applications (PROTEO), Université Laval, Québec, QC, Canada. nick.bisson@crchudequebec.ulaval.ca.|Department of Molecular Biology, Medical Biochemistry and Pathology, Université Laval, Québec, QC, Canada. nick.bisson@crchudequebec.ulaval.ca.
authors: Nicolas
authors: N
authors: Bisson
mesh_terms: D045744:Cell Line, Tumor; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D009369:Neoplasms / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D060066:Protein Interaction Maps
keywords: Mass spectrometry; Protein interactions; Sample preparation; Spin tip; Tryptic digestion