a 2022

From nano- to milimolar: dimerization dissociation constant determination

NÁPLAVOVÁ, Alexandra, Aneta KOZELEKOVÁ and Jozef HRITZ

Basic information

Original name

From nano- to milimolar: dimerization dissociation constant determination

Name (in English)

From nano- to milimolar: dimerization dissociation constant determination

Authors

NÁPLAVOVÁ, Alexandra, Aneta KOZELEKOVÁ and Jozef HRITZ

Edition

Chemistry towards Biology (CTB10), 2022

Other information

Type of outcome

Konferenční abstrakt

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

ISBN

978-80-971665-3-3

Keywords (in Czech)

14-3-3 protein; dimerizace; oligomerizace proteinů; 19F NMR

Keywords in English

14-3-3 protein; dimerization; protein oligomerization; 19F NMR
Změněno: 13/10/2022 15:27, Mgr. Alexandra Náplavová

Abstract

V originále

The self-association of proteins is the cornerstone of protein regulation, aiding proper functionality and interactome [1]. To properly understand the role of dimerization of protein, only the detection of such dimer is not sufficient, but a quantitative analysis is crucial. A parameter widely used to quantify the self-association is the dissociation constant KD [2]. The KD describes equilibrium between monomers and higher oligomers - in general, the lower the KD, the higher the propensity for oligomerization. However, KD of various proteins differs in several orders of magnitude, providing a challenge in its determination [3]. Here we showcase an array of biophysical methods for coverage of the whole relevant concentration range. For proteins with KD in nM region we optimised Förster resonance energy transfer assay [4]. In μM range an analytical size exclusion combined with multiple angle light scattering can be employed [3]. Finally, we used 19F Trp NMR for evaluation in higher concentrations. Moreover, we show application of such methods on the example of 14–3–3 proteins – cellular regulators connected to oncologic and neurodegenerative diseases [5,6].

Links

LM2018127, large research infrastructures
Name: Czech Infrastructure for Integrative Structural Biology (Acronym: CIISB)
Investor: Ministry of Education, Youth and Sports of the CR
LM2018127, large research infrastructures
Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)
Investor: Ministry of Education, Youth and Sports of the CR
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