An intermolecular FRET sensor detects the dynamics of T cell receptor clustering
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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.
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MA, Yuanqing, Elvis PANDZIC, Philip R. NICOVICH, Yui YAMAMOTO, Joanna KWIATEK, Sophie V. PAGEON, Aleš BENDA, Jérémie ROSSY, Katharina GAUS, 2017. An intermolecular FRET sensor detects the dynamics of T cell receptor clustering. In: Nature Communications. 2017, 8, 15100. eISSN 2041-1723. Available under: doi: 10.1038/ncomms15100BibTex
@article{Ma2017-04-28inter-42983, year={2017}, doi={10.1038/ncomms15100}, title={An intermolecular FRET sensor detects the dynamics of T cell receptor clustering}, volume={8}, journal={Nature Communications}, author={Ma, Yuanqing and Pandzic, Elvis and Nicovich, Philip R. and Yamamoto, Yui and Kwiatek, Joanna and Pageon, Sophie V. and Benda, Aleš and Rossy, Jérémie and Gaus, Katharina}, note={Article Number: 15100} }
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