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

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.

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

• Original language: English
• Type of outcome: Conference abstract
• Confidentiality degree: is not subject to a state or trade secret
• WWW: 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
• Tags: International impact

Abstract

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