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Phosphoprotein Enrichment for Protein Analysis in Motile T-Lymphocytes

  • Praseetha PrasannanEmail author
  • Rathi Saravanan
  • Navin Kumar Verma
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1930)

Abstract

Protein phosphorylation plays a key role in intracellular signal transduction and regulates diverse cellular functions. This posttranslational modification of proteins occurs dynamically and reversibly and only a small fraction of the total proteins is phosphorylated at any given time depending on the cell types and their functioning. Thus, a relatively low abundance of phosphorylated proteins is present in specific cells under certain conditions and hence it becomes problematic to detect these proteins and their analysis. In particular, phosphoproteomic analysis of rapidly migrating T-lymphocytes is always challenging. In order to analyze phosphoproteins in motile T-cells using techniques such as polyacrylamide gel electrophoresis and mass spectrometry, it is often important to enrich the phosphorylated forms in the cellular lysates. In this chapter, we describe a simple method to enrich phosphoproteins that can be used for protein analysis in motile T-cells.

Key words

Protein phosphorylation Phosphoproteomics T-cell migration Mass spectrometry 

Notes

Acknowledgments

This work was supported by the Lee Kong Chian School of Medicine, Nanyang Technological University Singapore Start-Up Grant to N.K.V. and the Singapore Ministry of Education (MOE) under its Singapore MOE Academic Research Fund (AcRF) Tier 2 Grant (MOE2017-T2-2-004).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Praseetha Prasannan
    • 1
    Email author
  • Rathi Saravanan
    • 1
  • Navin Kumar Verma
    • 2
  1. 1.Lymphocyte Signalling Research Laboratory, Lee Kong Chian School of MedicineNanyang Technological University SingaporeSingaporeSingapore
  2. 2.Lee Kong Chian School of MedicineNanyang Technological University SingaporeSingaporeSingapore

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