High-order fluorescence fluctuation analysis of model protein clusters.

The technique of high-order fluorescence fluctuation autocorrelation for detecting and characterizing protein oligomers was applied to solutions containing two fluorescent proteins in which the more fluorescent proteins were analogues for clusters of the less fluorescent ones. The results show that the model protein clusters can be detected for average numbers of observed subunits (free monomers plus monomers in oligomers) equal to 10-100 and for relative fluorescent yields that correspond to oligomers as small as trimers. High-order fluorescent fluctuation analysis may therefore be applicable to cell surface receptor clusters in natural or model membranes.