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Profiling Activity of Cellular Kinases in Migrating T-Cells

  • Chandra Sekhar Chirumamilla
  • Mobashar Hussain Urf Turabe FazilEmail author
  • Claudina Perez-Novo
  • Savithri Rangarajan
  • Rik de Wijn
  • Padma Ramireddy
  • Navin Kumar Verma
  • Wim Vanden Berghe
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1930)

Abstract

T-Lymphocyte kinases are important checkpoints that control T-cell motility by regulating a diverse range of signal transduction pathways. The distinct configuration of kinase events in T-cell could be used to fingerprint the status of T-cells. However, only small fraction human kinases have been characterized so far and little is known about the dynamics of the kinome in motile T-cells. Although several direct and indirect strategies exist to characterize cellular kinase activities, such as RNA interference, antibody arrays, enzyme kinetics, and mass spectrometry, this chapter focuses on an alternative multiplex phosphopeptide array-based methodology, which allows the kinome-wide identification of hyper-activated kinases involved in the regulation of T-cell migration.

Key words

Peptide microarray LFA-1 ICAM-1 Kinome analysis Kinase T-cell 

Notes

Acknowledgements

The authors acknowledge funding support from the Lee Kong Chian School of Medicine, Nanyang Technological University Singapore Start-Up Grant to N.K.V. and the Ministry of Education (MOE) Singapore under its Singapore MOE Academic Research Fund (AcRF) Tier 1 (2014-T1-001-141) and MOE-AcRF Tier 2 (MOE2017-T2-2-004) grants and Hercules Grant, U Antwerp to W.V.B.

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Copyright information

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

Authors and Affiliations

  • Chandra Sekhar Chirumamilla
    • 1
  • Mobashar Hussain Urf Turabe Fazil
    • 2
    Email author
  • Claudina Perez-Novo
    • 1
  • Savithri Rangarajan
    • 3
  • Rik de Wijn
    • 3
  • Padma Ramireddy
    • 1
  • Navin Kumar Verma
    • 4
  • Wim Vanden Berghe
    • 1
  1. 1.Laboratory of Protein Chemistry, Proteomics and Epigenetic Signalling (PPES), Department of Biomedical SciencesUniversity of Antwerp (UA)AntwerpenBelgium
  2. 2.Lymphocyte Signalling Research Laboratory, Lee Kong Chain School of MedicineNanyang Technological University SingaporeSingaporeSingapore
  3. 3.PamGene International B.V.5200 BJ ’s-HertogenboschThe Netherlands
  4. 4.Lee Kong Chain School of MedicineNanyang Technological University SingaporeSingaporeSingapore

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