Metallothionein dimerization evidenced by QD-based Förster resonance energy transfer and capillary electrophoresis

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216305%3A26310%2F21%3APU138262" target="_blank" >RIV/00216305:26310/21:PU138262 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/62156489:43210/21:43918913 RIV/00216224:14310/21:00121520

Výsledek na webu

<a href="https://www.sciencedirect.com/science/article/pii/S0141813020352466?via=ihub" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0141813020352466?via=ihub</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.ijbiomac.2020.12.105" target="_blank" >10.1016/j.ijbiomac.2020.12.105</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Metallothionein dimerization evidenced by QD-based Förster resonance energy transfer and capillary electrophoresis

Popis výsledku v původním jazyce

Herein, we report a new simple and easy-to-use approach for the characterization of protein oligomerization based on fluorescence resonance energy transfer (FRET) and capillary electrophoresis with LED-induced detection. The FRET pair consisted of quantum dots (QDs) used as an emission tunable donor (emission wavelength of 450 nm) and a cyanine dye (Cy3), providing optimal optical properties as an acceptor. Nonoxidative dimerization of mammalian metallothionein (MT) was investigated using the donor and acceptor covalently conjugated to MT. The main functions of MTs within an organism include the transport and storage of essential metal ions and detoxification of toxic ions. Upon storage under aerobic conditions, MTs form dimers (as well as higher oligomers), which may play an essential role as mediators in oxidoreduction signaling pathways. Due to metal bridging by Cd2+ ions between molecules of metallothionein, the QDs and Cy3 were close enough, enabling a FRET signal. The FRET efficiency was calculated to be in the range of 11–77%. The formation of MT dimers in the presence of Cd2+ ions was confirmed by MALDI-MS analyses. Finally, the process of oligomerization resulting in FRET was monitored by CE, and oligomerization of MT was confirmed.

Název v anglickém jazyce

Metallothionein dimerization evidenced by QD-based Förster resonance energy transfer and capillary electrophoresis

Popis výsledku anglicky

Herein, we report a new simple and easy-to-use approach for the characterization of protein oligomerization based on fluorescence resonance energy transfer (FRET) and capillary electrophoresis with LED-induced detection. The FRET pair consisted of quantum dots (QDs) used as an emission tunable donor (emission wavelength of 450 nm) and a cyanine dye (Cy3), providing optimal optical properties as an acceptor. Nonoxidative dimerization of mammalian metallothionein (MT) was investigated using the donor and acceptor covalently conjugated to MT. The main functions of MTs within an organism include the transport and storage of essential metal ions and detoxification of toxic ions. Upon storage under aerobic conditions, MTs form dimers (as well as higher oligomers), which may play an essential role as mediators in oxidoreduction signaling pathways. Due to metal bridging by Cd2+ ions between molecules of metallothionein, the QDs and Cy3 were close enough, enabling a FRET signal. The FRET efficiency was calculated to be in the range of 11–77%. The formation of MT dimers in the presence of Cd2+ ions was confirmed by MALDI-MS analyses. Finally, the process of oligomerization resulting in FRET was monitored by CE, and oligomerization of MT was confirmed.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>S - Specificky vyzkum na vysokych skolach

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES

ISSN

0141-8130

e-ISSN

1879-0003

Svazek periodika

170

Číslo periodika v rámci svazku

2021

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

8

Strana od-do

53-60

Kód UT WoS článku

000613921400006

EID výsledku v databázi Scopus

2-s2.0-85098131107