Detailed Information on Publication Record
2022
Quantitation of Human 14-3-3ζ Dimerization and the Effect of Phosphorylation on Dimer-monomer Equilibria
TROŠANOVÁ, Zuzana, Petr LOUŠA, Aneta KOZELEKOVÁ, Tomáš BROM, Norbert GAŠPARIK et. al.Basic information
Original name
Quantitation of Human 14-3-3ζ Dimerization and the Effect of Phosphorylation on Dimer-monomer Equilibria
Authors
TROŠANOVÁ, Zuzana (703 Slovakia, belonging to the institution), Petr LOUŠA (203 Czech Republic, belonging to the institution), Aneta KOZELEKOVÁ (203 Czech Republic, belonging to the institution), Tomáš BROM (203 Czech Republic, belonging to the institution), Norbert GAŠPARIK (703 Slovakia, belonging to the institution), Ján TUNGLI (703 Slovakia, belonging to the institution), Veronika WEISOVÁ (703 Slovakia, belonging to the institution), Erik ŽUPA (703 Slovakia, belonging to the institution), Gabriel ŽOLDÁK and Jozef HRITZ (703 Slovakia, guarantor, belonging to the institution)
Edition
Journal of Molecular Biology, Elsevier, 2022, 0022-2836
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10608 Biochemistry and molecular biology
Country of publisher
United Kingdom of Great Britain and Northern Ireland
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
Impact factor
Impact factor: 5.600
RIV identification code
RIV/00216224:14740/22:00125485
Organization unit
Central European Institute of Technology
UT WoS
000791750700015
Keywords in English
14-3-3; phosphorylation; dimerization; dissociation constant; FRET; NMR
Tags
International impact, Reviewed
Změněno: 9/10/2024 13:16, Ing. Martina Blahová
Abstract
V originále
14-3-3 proteins are universal regulatory proteins and their function depends on their oligomeric form which may alter between the monomeric, homodimeric and heterodimeric states. The populations of individual oligomeric forms are controlled by Kd values of the dimer–monomer equilibria between the involved isoforms. This complex picture is extended by post-translational modifications, e.g. phosphorylation. In this work, we describe the equilibria between monomers, homo- and heterodimers of the 14-3-3ζ isoform in the unmodified and phosphorylated form. To cover a wide range of dimerization affinities, we combined solution NMR, microscale thermophoresis, native PAGE, and a set of novel fluorescence assays. Using a FRET based assay, we also determined the kinetic parameters of dimerization. We found that phosphorylation of 14-3-3ζ at Ser58 increases its homodimeric Kd value by 6 orders of magnitude. The presented assays allow to efficiently monitor 14-3-3ζ dimerization as a function of external factors, such as temperature, salt concentration, and client protein binding. For instance, we obtained values of both transient and equilibrium thermodynamic constants for the dimerization, and observed a substantial decrease of 14-3-3ζ dimer dissociation rate upon binding to the doubly phosphorylated regulatory domain of tyrosine hydroxylase. In summary, our work provides a conceptual framework to characterise the isoform exchanges of homo- and heterodimers which can significantly deepen our knowledge about the regulatory function of 14-3-3 proteins.
Links
| GF20-05789L, research and development project |
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| LTAUSA18168, research and development project |
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| 7AMB16DE005, research and development project |
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| 90043, large research infrastructures |
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