19F Trp labelling as a selective probe into the dimeric
interface

a 2022

19F Trp labelling as a selective probe into the dimeric interface

NÁPLAVOVÁ, Alexandra, Aneta KOZELEKOVÁ, Norbert GAŠPARIK and Jozef HRITZ

Basic information

Original name

19F Trp labelling as a selective probe into the dimeric interface

Authors

NÁPLAVOVÁ, Alexandra, Aneta KOZELEKOVÁ, Norbert GAŠPARIK and Jozef HRITZ

Edition

Instruct Biennial Structural Biology Conference 2022, 2022

Other information

Language

English

Type of outcome

Konferenční abstrakt

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Organization unit

Faculty of Science

Keywords (in Czech)

14-3-3 protein; 19F NMR; 19F Trp značení

Keywords in English

14-3-3 protein; 19F NMR; 19F Trp labelling

Abstract

V originále

The self-association of proteins is a fundamental mechanism in cell. The oligomerization offers a way of protein regulation, often broadening their functionality [1]. One of the rarest amino acids tryptophan (Trp) has a unique role in protein self-association [2]. It has been previously described that Trp is commonly present in so called dimerization hot spots partaking in dimer formation [3,4]. A method that could selectively focus on Trp could therefore offer a unique probe into protein dimerization. Nuclear magnetic resonance is a powerful tool in structural biology. Traditional double 13C, 15N labelling can be complemented by the 19F labelling which is a simple and straight-forward method [5]. The biggest advantage of 19F labelling is high signal intensity and a selective labelling of only chosen amino acid (usually Trp, Tyr or Phe), leading to less convoluted spectra without background noise. For these reasons, the 19F labelling may present a technique especially useful for determination of parameters connected to protein dimerization. In this work, we explore application of 19F Trp labelling on example of 14-3-3 proteins. The eukaryotic 14-3-3 proteins are important regulators involved in number of processes. Their dimeric form is crucial for proper function and the mechanism of dimer-ligand interaction has been thoroughly described [6]. Upon study of post-translational modifications, it has been discovered that phosphorylation of 14-3-3 at Ser58 located at dimeric interface leads to monomerization [7]. Intriguingly, residue neighbouring Ser58 is Trp59. We are thus comparing 19F Trp labelled dimeric 14-3-3 wild type and monomeric 14-3-3 phosphorylated at Ser58 and showing the possibilities that such labelling offers for study of dimerization.

Links

LTAUSA18168, research and development project

Name: Selektivní NMR značení jako nástroj pro charakterizaci proteinových komplexů zapojených do neurodegenerativních onemocnění

Investor: Ministry of Education, Youth and Sports of the CR, Selective NMR labelling as a tool for characterization of protein complexes involved in neurodegenerative diseases., INTER-ACTION

MUNI/C/0091/2022, interní kód MU

Name: Structural and interaction properties of 14-3-3 proteins in the monomeric state.

Investor: Masaryk University, Excellent diploma thesis

Citovat

NÁPLAVOVÁ, Alexandra, Aneta KOZELEKOVÁ, Norbert GAŠPARIK and Jozef HRITZ. 19F Trp labelling as a selective probe into the dimeric interface. In Instruct Biennial Structural Biology Conference 2022. 2022.

@proceedings{1855579,
   author = {Náplavová, Alexandra and Kozeleková, Aneta and Gašparik, Norbert and Hritz, Jozef},
   booktitle = {Instruct Biennial Structural Biology Conference 2022},
   keywords = {14-3-3 protein; 19F NMR; 19F Trp labelling},
   language = {eng},
   title = {19F Trp labelling as a selective probe into the dimeric interface},
   url = {https://instruct-eric.org/download/?file=Biennial_2022_Abstract_Booklet_2022.pdf},
   year = {2022}
}

TY  - CONF
ID  - 1855579
AU  - Náplavová, Alexandra - Kozeleková, Aneta - Gašparik, Norbert - Hritz, Jozef
PY  - 2022
TI  - 19F Trp labelling as a selective probe into the dimeric interface
KW  - 14-3-3 protein
KW  - 19F NMR
KW  - 19F Trp labelling
UR  - https://instruct-eric.org/download/?file=Biennial_2022_Abstract_Booklet_2022.pdf
N2  - The self-association of proteins is a fundamental mechanism in cell. The oligomerization offers a way of protein regulation, often broadening their functionality [1]. One of the rarest amino acids tryptophan (Trp) has a unique role in protein self-association [2]. It has been previously described that Trp is commonly present in so called dimerization hot spots partaking in dimer formation [3,4]. A method that could selectively focus on Trp could therefore offer a unique probe into protein dimerization. Nuclear magnetic resonance is a powerful tool in structural biology. Traditional double 13C, 15N labelling can be complemented by the 19F labelling which is a simple and straight-forward method [5]. The biggest advantage of 19F labelling is high signal intensity and a selective labelling of only chosen amino acid (usually Trp, Tyr or Phe), leading to less convoluted spectra without background noise. For these reasons, the 19F labelling may present a technique especially useful for determination of parameters connected to protein dimerization. In this work, we explore application of 19F Trp labelling on example of 14-3-3 proteins. The eukaryotic 14-3-3 proteins are important regulators involved in number of processes. Their dimeric form is crucial for proper function and the mechanism of dimer-ligand interaction has been thoroughly described [6]. Upon study of post-translational modifications, it has been discovered that phosphorylation of 14-3-3 at Ser58 located at dimeric interface leads to monomerization [7]. Intriguingly, residue neighbouring Ser58 is Trp59. We are thus comparing 19F Trp labelled dimeric 14-3-3 wild type and monomeric 14-3-3 phosphorylated at Ser58 and showing the possibilities that such labelling offers for study of dimerization.
ER  -

NÁPLAVOVÁ, Alexandra, Aneta KOZELEKOVÁ, Norbert GAŠPARIK and Jozef HRITZ. 19F Trp labelling as a selective probe into the dimeric interface. In \textit{Instruct Biennial Structural Biology Conference 2022}. 2022.

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