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Further insights into the metal ion binding abilities and the metalation pathway of a plant metallothionein from Musa acuminata

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Abstract

The superfamily of metallothioneins (MTs) combines a diverse group of metalloproteins, sharing the characteristics of rather low molecular weight and high cysteine content. The latter provides MTs with the capability to coordinate thiophilic metal ions, in particular those with a d 10 electron configuration. The sub-family of plant MT3 proteins is only poorly characterized and there is a complete lack of three-dimensional structure information. Building upon our previous results on the Musa acuminata MT3 (musMT3) protein, the focus of the present work is to understand the metal cluster formation process, the role of the single histidine residue present in musMT3, and the metal ion binding affinity. We concentrate our efforts on the coordination of ZnII and CdII ions, using CoII as a spectroscopic probe for ZnII binding. The overall protein-fold is analysed with a combination of limited proteolytic digestion, mass spectrometry, and dynamic light scattering. Histidine coordination of metal ions is probed with extended X-ray absorption fine structure spectroscopy and CoII titration experiments. Initial experiments with isothermal titration calorimetry provide insights into the thermodynamics of metal ion binding.

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Acknowledgements

The authors acknowledge funding from the Swiss National Science Foundation (E.F.) and the National Institutes of Health (V.L.P., ES012236; A.D. and J.E.P.-H., GM 38047) for support of this research. Synchrotron measurements were made at the Stanford Synchrotron Radiation Laboratory, which is supported by the NIH Research Resource Program and the US Department of Energy.

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Correspondence to Eva Freisinger.

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This work is dedicated to Professor Dr. Dr. h.c. Helmut Sigel with the most heartfelt wishes for his 80th birthday and many, many years to come.

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Cabral, A.C.S., Jakovleska, J., Deb, A. et al. Further insights into the metal ion binding abilities and the metalation pathway of a plant metallothionein from Musa acuminata . J Biol Inorg Chem 23, 91–107 (2018). https://doi.org/10.1007/s00775-017-1513-9

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