title: Phosphoproteomic Analysis of Cell-Based Resistance to BRAF Inhibitor Therapy in Melanoma.
abstract: The treatment of melanoma by targeted inhibition of the mutated kinase BRAF with small molecules only temporarily suppresses metastatic disease. In the face of chemical inhibition tumor plasticity, both innate and adaptive, promotes survival through the biochemical and genetic reconfiguration of cellular pathways that can engage proliferative and migratory systems. To investigate this process, high-resolution mass spectrometry was used to characterize the phosphoproteome of this transition in vitro. A simple and accurate, label-free quantitative method was used to localize and quantitate thousands of phosphorylation events. We also correlated changes in the phosphoproteome with the proteome to more accurately determine changes in the activity of regulatory kinases determined by kinase landscape profiling. The abundance of phosphopeptides with sites that function in cytoskeletal regulation, GTP/GDP exchange, protein kinase C, IGF signaling, and melanosome maturation were highly divergent after transition to a drug resistant phenotype.
authors.affiliation: Australian Proteome Analysis Facility, Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, NSW , Australia.
authors.forename: Robert
authors.initials: R
authors.lastname: Parker
authors.affiliation: Cancer Immunology Group, Olivia Newton-John Cancer Research Institute, Ludwig Institute for Cancer Research, School of Cancer Medicine, La Trobe University , Heidelberg, VIC , Australia.
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Vella
authors.affiliation: Australian Proteome Analysis Facility, Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, NSW , Australia.
authors.forename: Dylan
authors.initials: D
authors.lastname: Xavier
authors.affiliation: Australian Proteome Analysis Facility, Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, NSW , Australia.
authors.forename: Ardeshir
authors.initials: A
authors.lastname: Amirkhani
authors.affiliation: NHS Trust Southport and Ormskirk General Hospital , Ormskirk , UK.
authors.forename: Jimmy
authors.initials: J
authors.lastname: Parker
authors.affiliation: Cancer Immunology Group, Olivia Newton-John Cancer Research Institute, Ludwig Institute for Cancer Research, School of Cancer Medicine, La Trobe University , Heidelberg, VIC , Australia.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Cebon
authors.affiliation: Australian Proteome Analysis Facility, Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, NSW , Australia.
authors.forename: Mark P
authors.initials: MP
authors.lastname: Molloy
keywords: BRAF; drug resistance; kinases; label-free quantitation; mass spectrometry; phosphoproteomics; vemurafenib