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

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.

journal_name

Nat Commun

journal_title

Nature communications

authors

Ma Y,Pandzic E,Nicovich PR,Yamamoto Y,Kwiatek J,Pageon SV,Benda A,Rossy J,Gaus K

doi

10.1038/ncomms15100

subject

Has Abstract

pub_date

2017-04-28 00:00:00

pages

15100

issn

2041-1723

pii

ncomms15100

journal_volume

8

pub_type

杂志文章

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