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

作者：Ma, Yuanqing; Pandzic, Elvis; Nicovich, Philip R.; Yamamoto, Yui; Kwiatek, Joanna; Pageon, Sophie V.; Benda, Ales; Rossy, Jeremie; Gaus, Katharina^*

来源：Nature Communications, 2017, 8(1): 15100.

DOI：10.1038/ncomms15100

• IMMUNOLOGICAL SYNAPSE
• LIVING CELLS
• HOMO-FRET
• ACTIVATION
• MICROCLUSTERS
• MEMBRANE
• MICROSCOPY
• AGONIST
• DOMAIN

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 Forster 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.

• 出版日期2017-4-28