Examining Cellular Responses to Kinase Drug Inhibition Through Phosphoproteome Mapping of Substrates

Daniel Bucio-Noble; Crystal Semaan; Mark P Molloy

doi:10.1007/978-1-4939-8891-4_8

Examining Cellular Responses to Kinase Drug Inhibition Through Phosphoproteome Mapping of Substrates

Methods Mol Biol. 2019:1888:141-152. doi: 10.1007/978-1-4939-8891-4_8.

Authors

Daniel Bucio-Noble¹, Crystal Semaan¹, Mark P Molloy^{2

3

4}

Affiliations

¹ Department of Molecular Sciences, ARC Training Centre for Molecular Technology in the Food Industry, Macquarie University, Sydney, Australia.
² Department of Molecular Sciences, ARC Training Centre for Molecular Technology in the Food Industry, Macquarie University, Sydney, Australia. m.molloy@sydney.edu.au.
³ Australian Proteome Analysis Facility, Macquarie University, Sydney, Australia. m.molloy@sydney.edu.au.
⁴ Bowel Cancer and Biomarker Research, Kolling Institute, The University of Sydney, Sydney, Australia. m.molloy@sydney.edu.au.

PMID: 30519945
DOI: 10.1007/978-1-4939-8891-4_8

Abstract

This chapter provides a detailed description for mass spectrometry-based phosphoproteomics analysis. We describe sample preparation, phosphopeptide enrichment, mass spectrometry acquisition, label-free data analysis and statistical analysis. This technique can be employed to characterize cell signaling networks, and is particularly useful to monitor cellular responses to kinase drug inhibitors.

Keywords: Label-free; Mass spectrometry; Phosphopeptide enrichment; Phosphoproteomics.

MeSH terms

Chromatography, Liquid
Drug Discovery* / methods
Enzyme Inhibitors / pharmacology*
Humans
Mass Spectrometry
Phosphopeptides / metabolism
Phosphoproteins / metabolism*
Phosphotransferases / antagonists & inhibitors*
Proteomics* / methods

Substances

Enzyme Inhibitors
Phosphopeptides
Phosphoproteins
Phosphotransferases