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PAVELICOVÁ, K. POMPEIANO VANÍČKOVÁ, L. HADDAD, Y. NEJDL, L. ZÍTKA, J. KOČIOVÁ, S. MRAVEC, F. VACULOVIČ, T. MACKA, M. VACULOVIČOVÁ, M. ADAM, V.
Original Title
Metallothionein dimerization evidenced by QD-based Förster resonance energy transfer and capillary electrophoresis
Type
journal article in Web of Science
Language
English
Original Abstract
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.
Keywords
FRET;Quantum dots;Oligomerization;Capillary electrophoresis
Authors
PAVELICOVÁ, K.; POMPEIANO VANÍČKOVÁ, L.; HADDAD, Y.; NEJDL, L.; ZÍTKA, J.; KOČIOVÁ, S.; MRAVEC, F.; VACULOVIČ, T.; MACKA, M.; VACULOVIČOVÁ, M.; ADAM, V.
Released
15. 2. 2021
Publisher
Elsevier
Location
Nizozemsko
ISBN
0141-8130
Periodical
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Year of study
170
Number
1
State
Kingdom of the Netherlands
Pages from
53
Pages to
60
Pages count
8
URL
https://www.sciencedirect.com/science/article/pii/S0141813020352466
Full text in the Digital Library
http://hdl.handle.net/11012/208291
BibTex
@article{BUT167427, author="Kristýna {Pavelicová} and Lucie {Pompeiano Vaníčková} and Yazan Abdulmajeed Eyadh {Haddad} and Lukáš {Nejdl} and Jan {Zítka} and Silvia {Kočiová} and Filip {Mravec} and Tomáš {Vaculovič} and Mirek {Macka} and Markéta {Vaculovičová} and Vojtěch {Adam}", title="Metallothionein dimerization evidenced by QD-based Förster resonance energy transfer and capillary electrophoresis", journal="INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES", year="2021", volume="170", number="1", pages="53--60", doi="10.1016/j.ijbiomac.2020.12.105", issn="0141-8130", url="https://www.sciencedirect.com/science/article/pii/S0141813020352466" }