14-3-3 protein isoforms: mutual dimerization propensities screening kit

ŠIMEK, Jan, Kateřina KRÁLOVÁ and Jozef HRITZ. 14-3-3 protein isoforms: mutual dimerization propensities screening kit. 2022.

Basic information

• Original name: 14-3-3 protein isoforms: mutual dimerization propensities screening kit
• Authors: ŠIMEK, Jan, Kateřina KRÁLOVÁ and Jozef HRITZ.
• Edition: 2022.

Other information

• Type of outcome: Conference abstract
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• Keywords (in Czech): 14-3-3 protein; homodimerizace, heterodimerizace, fluorescence, FRET, kinetika, disociační konstanta
• Keywords in English: 14-3-3 protein; homodimerization, heterodimerization, fluorescence, FRET, kinetics, dissociation constant

Abstract

14-3-3 is a group of eukaryotic regulatory proteins, well known for its irreplaceable role in regulation of cell cycle and metabolism. 14-3-3 family involves 7 isoforms of this protein: ε, γ, ζ, β, θ, η and σ.[1] These isoforms differ in their sequences, expression levels in various tissues and interactomes, but their structures and general biophysical properties are similar. Function of 14-3-3 protein is determined by its dimeric character. [2] It was observed that 14-3-3 proteins are capable of both homoand heterodimerization. Due to different sequences of dimeric interfaces, monomeric subunits of different isoforms have different mutual affinities.[3], [4] Target of this work is to compare dimerization propensities of all possible pairs of 14-3-3 monomers. In our research group, the dimerization of the ζ isoform was deeply studied, using FRET fluorescence assay in 1.5 ml cuvette, and evaluated applying complex mathematical model. This experiment provides quantitative description of dimerization, but can be performed only for one dimerization pair and is relatively time consuming. [5] Therefore, in this study, fluorimetric screening on 96 well plate was developed to describe mutual affinities of all 14-3-3 isoforms. Main output is rough information about which pairs could not form dimers, and which are candidates for further studies.

Links

• GF20-05789L, research and development project: Name: Charakterizace přirozeně neuspořádaných proteinů

Investor: Czech Science Foundation

• LM2018127, research and development project: Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR