Quantification of integrin receptor agonism by fluorescence lifetime imaging.

ABSTRACT: Both spatiotemporal analyses of adhesion signalling and the development of pharmacological inhibitors of integrin receptors currently suffer from the lack of an assay to measure integrin-effector binding and the response of these interactions to antagonists. Indeed, anti-integrin compounds have failed in the clinic because of secondary side effects resulting from agonistic activity. Here, we have expressed integrin-GFP and effector-mRFP pairs in living cells and quantified their association using fluorescence lifetime imaging microscopy (FLIM) to measure fluorescence resonance energy transfer (FRET). Association of talin with beta1 integrin and paxillin with alpha4 integrin was dependent on both the ligand and receptor activation state, and was sensitive to inhibition with small molecule R

SUBMITTER: Parsons M 

PROVIDER: S-EPMC3328206 | biostudies-literature | 2008 Feb

REPOSITORIES: biostudies-literature