The 14-3 -3 protein family plays a central role in signal transduction by recognizing phosphorylated serine/threonine sequence motifs in client proteins and by modulating their function. These sequence motifs are bound by the two binding sites of a 14-3-3 protein, which are only present in a dimeric structure. Phosphorylation of S58 within the 14-3-3 homodimer triggers dissociation of the monomeric subunits, which may impair their function. Thus, while having appreciated importance of the integrity of dimeric structure for a proper biological function, no quantitative analytical tools for dimer-monomer equilibrium have been developed until now.
In the current proposal, an international, interdisciplinary team of researchers suggests to develop an efficient fluorescence-based approach for studying equilibrium and kinetics of quaternary structure formation of members of the 14-3-3 families. The 14-3-3zeta protein will be purified, specifically labeled by fluorescent dyes and used for FRET experiments, microscale thermophoresis, NMR as well as being investigated by computational methods.

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