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

a 2022

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

ŠIMEK, Jan, Kateřina KRÁLOVÁ and Jozef HRITZ

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

Konferenční abstrakt

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

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

Změněno: 17/4/2023 13:33, Mgr. Jan Šimek

Abstract

V originále

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, Partner Agency (Austria)

LM2018127, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

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

Citovat

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

@proceedings{2275859,
   author = {Šimek, Jan and Králová, Kateřina and Hritz, Jozef},
   keywords = {14-3-3 protein; homodimerization, heterodimerization, fluorescence, FRET, kinetics, dissociation constant},
   title = {14-3-3 protein isoforms: mutual dimerization propensities screening kit},
   url = {https://instruct-eric.org/download/?file=Biennial_2022_Abstract_Booklet_2022.pdf&do=do},
   year = {2022}
}

TY  - CONF
ID  - 2275859
AU  - Šimek, Jan - Králová, Kateřina - Hritz, Jozef
PY  - 2022
TI  - 14-3-3 protein isoforms: mutual dimerization propensities screening kit
KW  - 14-3-3 protein
KW  - homodimerization, heterodimerization, fluorescence, FRET, kinetics, dissociation constant
UR  - https://instruct-eric.org/download/?file=Biennial_2022_Abstract_Booklet_2022.pdf&do=do
N2  - 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.
ER  -

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

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