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 a Jozef HRITZ

Základní údaje

Originální název

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

Autoři

ŠIMEK, Jan; Kateřina KRÁLOVÁ a Jozef HRITZ

Vydání

2022

Další údaje

Typ výsledku

Konferenční abstrakt

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Klíčová slova česky

14-3-3 protein; homodimerizace, heterodimerizace, fluorescence, FRET, kinetika, disociační konstanta

Klíčová slova anglicky

14-3-3 protein; homodimerization, heterodimerization, fluorescence, FRET, kinetics, dissociation constant

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

Anotace

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.

Návaznosti

GF20-05789L, projekt VaV

Název: Charakterizace přirozeně neuspořádaných proteinů

Investor: Grantová agentura ČR, Characterization of intrinsically disordered proteins, Partnerská agentura (Rakousko)

LM2018127, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii (Akronym: CIISB)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Czech Infrastructure for Integrative Structural Biology

Citovat

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

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