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Spatial and Temporal Control of T Cell Activation Using a Photoactivatable Agonist
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Distinct Mechanisms Regulate Lck Spatial Organization in Activated T Cells.

Natasha Kapoor-Kaushik1, Elizabeth Hinde2, Ewoud B Compeer1

  • 1EMBL Australia Node in Single Molecule Science, School of Medical Science, University of New South Wales , Sydney, NSW , Australia.

Frontiers in Immunology
|March 26, 2016
PubMed
Summary

The kinase Lck

Keywords:
LckT cell signalingassembly of signaling complexesimage correlation spectroscopymembrane organizationsuper-resolution fluorescence microscopy

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Area of Science:

  • Immunology
  • Cell Biology
  • Biochemistry

Background:

  • T cell receptor (TCR) phosphorylation by Lck kinase initiates T cell signaling.
  • Lck's plasma membrane distribution, conformation, activity, and interactions regulate TCR phosphorylation efficiency.

Purpose of the Study:

  • To investigate the mechanisms governing Lck clustering and its impact on T cell activation.
  • To elucidate the roles of Lck conformation and phosphorylation in spatial organization.

Main Methods:

  • Utilized cross-correlation raster image correlation spectroscopy (2D-ccRICS) and photoactivated localization microscopy (PALM).
  • Investigated Lck clustering mechanisms, including intrinsic (conformation-dependent) and extrinsic (phosphorylation-dependent) pathways.

Main Results:

  • Identified two distinct Lck clustering mechanisms: one intrinsic to Lck's open conformation and another extrinsic, regulated by tyrosine 192 phosphorylation.
  • Observed differential effects of Zap70 and Lat absence on these clustering mechanisms.
  • Found that TSAd binding promotes Lck diffusion upon tyrosine 192 phosphorylation.

Conclusions:

  • Lck's open conformation drives aggregation, while downstream signaling events modulate its spatial organization.
  • These findings provide insights into the regulation of early T cell signaling events.