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

Citation

Pavelicova, K and Vanickova, L and Haddad, Y and Nejdl, L and Zitka, J and Kociova, S and Mravec, F and Vaculovic, T and Macka, M and Vaculovicova, M and Adam, V, Metallothionein dimerization evidenced by QD-based Forster resonance energy transfer and capillary electrophoresis, International Journal of Biological Macromolecules, 170 pp. 53-60. ISSN 0141-8130 (2021) [Refereed Article]


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DOI: doi:10.1016/j.ijbiomac.2020.12.105

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.

Item Details

Item Type:Refereed Article
Keywords:FRET, quantum dots, oligomerization, capillary electrophoresis
Research Division:Chemical Sciences
Research Group:Analytical chemistry
Research Field:Separation science
Objective Division:Expanding Knowledge
Objective Group:Expanding knowledge
Objective Field:Expanding knowledge in the chemical sciences
UTAS Author:Macka, M (Professor Mirek Macka)
ID Code:151801
Year Published:2021
Deposited By:Plant Science
Deposited On:2022-08-04
Last Modified:2022-08-04
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