Skip to main content

Detailed binding scheme and structural determination of the 14-3-3ζ in complex with a double phosphorylated human tyrosine hydroxylase 1

Final Report Summary - MODULATOR (Detailed binding scheme and structural determination of the 14-3-3ζ in complex with a double phosphorylated human tyrosine hydroxylase 1)

This early termination report covers 7 months period since 1st October 2013 - 30th April 2014.

Members of the 14-3-3 protein family are important modulators of several key signaling pathways that regulate critical biological activities, including cell cycle control, cell growth, proliferation, and apoptosis. Human tyrosine hydroxylase 1 (hTH1) activity is regulated by phosphorylation of its N-terminus and by an interaction with the modulator protein 14-3-3zeta. Human 14-3-3zeta isoforms form very stable homo-dimer.

In order to study the effect of monomerization of 14-3-3zeta protein, mutations at the dimer interface were designed and incorporated into the 14-3-zeta gene. Purity of the 14-3-3zeta mutants and wild type was checked by SDS-PAGE electrophoresis and MALDI mass spectrometry. NATIVE PAGE electrophoresis revealed that one of two tested 14-3-3zeta mutants exists almost exclusively in the monomeric state.
Other two high purity samples prepared within the time frame of the project were C-tail truncated construct of 14-3-3zeta and doubly phosphorylated peptide (1-50 region) of hTH1 (dp-hTH1-50). All samples will be further analyzed and used for detailed study of 14-3-3zeta protein monomerization.

The project web site: http://www.ceitec.eu/project-modulator/t1816