Quantifying protein-protein interactions of peripheral membrane proteins by fluorescence brightness analysis.

ABSTRACT: Fluorescently labeled proteins that are found both in the cytoplasm and at the plasma membrane, such as peripheral membrane proteins, create stratified fluorescent layers that present a challenging environment for brightness studies with fluorescence fluctuation spectroscopy. The geometry of each layer along with fluorescence and brightness contributions from adjacent layers generates a convoluted raw brightness that conceals the underlying brightness of each individual layer. Because the brightness at a layer establishes the oligomeric state of the fluorescently labeled protein at said layer, we developed a method that connects the experimental raw brightness with the physical brightness at each layered compartment. The technique determines the oligomerization in each compartment from an

SUBMITTER: Smith EM 

PROVIDER: S-EPMC4119281 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature