Phosphotyrosine Profiling Using SILAC

Keshava K Datta; Aditi Chatterjee; Harsha Gowda

doi:10.1007/978-1-0716-2863-8_9

Phosphotyrosine Profiling Using SILAC

Methods Mol Biol. 2023:2603:117-125. doi: 10.1007/978-1-0716-2863-8_9.

Authors

Keshava K Datta¹, Aditi Chatterjee², Harsha Gowda^{3

4

5}

Affiliations

¹ Department of Genetics and Computational Biology, QIMR Berghofer Medical Research Institute, Brisbane, QLD, Australia.
² Institute of Bioinformatics, Bangalore, India. aditixchatterjee@gmail.com.
³ Department of Genetics and Computational Biology, QIMR Berghofer Medical Research Institute, Brisbane, QLD, Australia. harshahc@gmail.com.
⁴ School of Biomedical Sciences, The University of Queensland, Brisbane, QLD, Australia. harshahc@gmail.com.
⁵ Faculty of Health, Queensland University of Technology, Brisbane, QLD, Australia. harshahc@gmail.com.

PMID: 36370274
DOI: 10.1007/978-1-0716-2863-8_9

Abstract

Tyrosine phosphorylation on proteins is an important posttranslational modification that regulates various processes in cells. Mass spectrometry-based phosphotyrosine profiling can reveal tyrosine kinase signaling activity in cells. Using quantitative proteomics strategies such as stable isotope labeling with amino acids in cell culture (SILAC) allows comparison of tyrosine kinase signaling activity across two to -three different conditions. In this book chapter, we discuss the reagents required and a step-by-step protocol to carry out phosphotyrosine profiling using SILAC.

Keywords: Kinases; Mass spectrometry; Phosphatases; Phosphopeptides; Phosphorylation; Proteomics; SILAC; Tyrosine.

MeSH terms

Isotope Labeling / methods
Phosphorylation
Phosphotyrosine / metabolism
Protein-Tyrosine Kinases* / metabolism
Proteomics* / methods

Substances

Phosphotyrosine
Protein-Tyrosine Kinases