An intermolecular FRET sensor detects the dynamics of T cell receptor clustering

Yuanqing Ma, Elvis Pandzic, Philip R. Nicovich, Yui Yamamoto, Joanna Kwiatek, Sophie V. Pageon, Aleš Benda, Jérémie Rossy and Katharina Gaus ()
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Yuanqing Ma: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales
Elvis Pandzic: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales
Philip R. Nicovich: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales
Yui Yamamoto: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales
Joanna Kwiatek: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales
Sophie V. Pageon: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales
Aleš Benda: Biomedical Imaging Facility, Lowy Cancer Research Centre, University of New South Wales
Jérémie Rossy: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales
Katharina Gaus: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales

Nature Communications, 2017, vol. 8, issue 1, 1-11

Abstract: Abstract Clustering of the T-cell receptor (TCR) is thought to initiate downstream signalling. However, the detection of protein clustering with high spatial and temporal resolution remains challenging. Here we establish a Förster resonance energy transfer (FRET) sensor, named CliF, which reports intermolecular associations of neighbouring proteins in live cells. A key advantage of the single-chain FRET sensor is that it can be combined with image correlation spectroscopy (ICS), single-particle tracking (SPT) and fluorescence lifetime imaging microscopy (FLIM). We test the sensor with a light-sensitive actuator that induces protein aggregation upon radiation with blue light. When applied to T cells, the sensor reveals that TCR triggering increases the number of dense TCR–CD3 clusters. Further, we find a correlation between cluster movement within the immunological synapse and cluster density. In conclusion, we develop a sensor that allows us to map the dynamics of protein clustering in live T cells.

Date: 2017
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DOI: 10.1038/ncomms15100

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