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

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Authors

PAVELICOVA Kristyna VANICKOVA Lucie HADDAD Yazan NEJDL Lukas ZITKA Jan KOCIOVA Silvia MRAVEC Filip VACULOVIČ Tomáš MACKA Mirek VACULOVICOVA Marketa ADAM Vojtech

Year of publication 2021
Type Article in Periodical
Magazine / Source International Journal of Biological Macromolecules
MU Faculty or unit

Faculty of Science

Citation
Web https://doi.org/10.1016/j.ijbiomac.2020.12.105
Doi http://dx.doi.org/10.1016/j.ijbiomac.2020.12.105
Keywords FRET; Quantum dots; Oligomerization; Capillary electrophoresis
Description 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. (C) 2020 Elsevier B.V. All rights reserved.
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