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Drosophila MTN: a metazoan copper-thionein related to fungal forms
M Valls1, R Bofill, N Romero-Isart
1Departament de Genètica, Facultat de Biologia, Av Diagonal 645, Universitat de Barcelona, E-08028, Barcelona, Spain.
Abstract:
Two Drosophila metallothioneins (MT) have been reported: MTN, a 40 residue peptide including 10 Cys, and MTO, a 43 residue peptide including 12 Cys. However, neither functional nor evolutionary analyses for either of the Drosophila MT are available. Here, heterologous expression of Mtn in Escherichia coli is reported. The metal binding abilities of the Cu- and Zn-MTN complexes conformed in vivo, as well as the features of the Cd- and Cu-aggregates produced by metal replacement in vitro, have been determined by atomic emission spectrometry, circular dichroism and electrospray ionization mass spectrometry. Primary structure relationships with other MT have been examined. The results indicate a close resemblance of MTN to fungal copper-thioneins.
Insights
This study reports the heterologous expression of Drosophila metallothionein (MTN) in E. coli. The findings reveal MTN
Area of Science:
- Biochemistry
- Molecular Biology
- Proteomics
Background:
- Two Drosophila metallothioneins (MTN, MTO) are known but lack functional and evolutionary analysis.
- Metallothioneins are cysteine-rich proteins involved in metal homeostasis and detoxification.
Purpose of the Study:
- To functionally characterize Drosophila metallothionein (MTN) through heterologous expression.
- To investigate the metal-binding properties and evolutionary relationships of MTN.
Main Methods:
- Heterologous expression of Drosophila MTN in Escherichia coli.
- Characterization using atomic emission spectrometry, circular dichroism, and electrospray ionization mass spectrometry.
- In vivo and in vitro metal-binding assays (Cu, Zn, Cd).
Main Results:
- Successful heterologous expression and purification of Drosophila MTN.
- Demonstrated in vivo metal-binding capabilities for copper (Cu) and zinc (Zn).
- Characterized in vitro Cd- and Cu-aggregate formation, showing structural similarities to fungal copper-thioneins.
Conclusions:
- Drosophila MTN exhibits functional metal-binding properties.
- MTN shares primary structural similarities with fungal copper-thioneins, suggesting conserved evolutionary roles.
- This work provides foundational functional data for Drosophila metallothioneins.