5 years ago

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

An intermolecular FRET sensor detects the dynamics of T cell receptor clustering
Elvis Pandzic, Joanna Kwiatek, Sophie V. Pageon, Jérémie Rossy, Katharina Gaus, Yuanqing Ma, Philip R. Nicovich, Aleš Benda, Yui Yamamoto
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.

Publisher URL: http://www.nature.com/articles/ncomms15100

DOI: 10.1038/ncomms15100

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