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Functional characterization of a type 3 metallolthionein isoform (OsMTI-3a) from rice.
Int J Biol Macromol. 2015 Feb; 73:154-9.IJ

Abstract

Metallothioneins (MTs) are low-molecular weight proteins with high Cys content and a high affinity for metals. Plant MTs are classified into four types based on the arrangement of Cys in their amino acid sequences. In the present study, the gene encoding OsMTI-3a, a type 3 MT found in rice, was cloned into pET41a vector. The resulting construct was transformed into the Escherichia coli strain Rosetta (DE3). Following the induction with isopropyl β-D-1-thiogalactopyranoside, the OsMTI-3a was expressed as glutathione-S-transferase (GST)-tagged fusion protein. In comparison to control strain, the cells expressing GST-OsMTI-3a accumulated more Cd(2+), Ni(2+) and Zn(2+) when they were grown in the medium containing CdCl2, NiCl2 or ZnSO4. The recombinant GST-OsMTI-3a was purified using affinity chromatography. The UV absorption spectra recorded after the reconstitution of the apo-protein with different metals confirmed that GST-OsMTI-3a was able to form complexes with Cd(2+), Ni(2+), and Zn(2+). The reaction of the protein-metal complexes with 5-5-dithiobis (2-nitrobenzoic) revealed that the order of affinity of GST-OsMTI-3a toward different metals was Ni(2+)≥Cd(2+)>Zn(2+)>Cu(2+).

Authors+Show Affiliations

Department of Agricultural Biotechnology, College of Agriculture, Isfahan University of Technology, Isfahan 84156-83111, Iran. Electronic address: a.shahpiri@cc.iut.ac.ir.Department of Agricultural Biotechnology, College of Agriculture, Isfahan University of Technology, Isfahan 84156-83111, Iran.Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan 81746-73441, Iran.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25449122

Citation

Shahpiri, Azar, et al. "Functional Characterization of a Type 3 Metallolthionein Isoform (OsMTI-3a) From Rice." International Journal of Biological Macromolecules, vol. 73, 2015, pp. 154-9.
Shahpiri A, Soleimanifard I, Asadollahi MA. Functional characterization of a type 3 metallolthionein isoform (OsMTI-3a) from rice. Int J Biol Macromol. 2015;73:154-9.
Shahpiri, A., Soleimanifard, I., & Asadollahi, M. A. (2015). Functional characterization of a type 3 metallolthionein isoform (OsMTI-3a) from rice. International Journal of Biological Macromolecules, 73, 154-9. https://doi.org/10.1016/j.ijbiomac.2014.10.067
Shahpiri A, Soleimanifard I, Asadollahi MA. Functional Characterization of a Type 3 Metallolthionein Isoform (OsMTI-3a) From Rice. Int J Biol Macromol. 2015;73:154-9. PubMed PMID: 25449122.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Functional characterization of a type 3 metallolthionein isoform (OsMTI-3a) from rice. AU - Shahpiri,Azar, AU - Soleimanifard,Iman, AU - Asadollahi,Mohammad Ali, Y1 - 2014/11/28/ PY - 2014/08/21/received PY - 2014/10/29/revised PY - 2014/10/30/accepted PY - 2014/12/3/entrez PY - 2014/12/3/pubmed PY - 2015/9/12/medline KW - Functional characterization KW - Heavy metals KW - Heterologous expression KW - Metallothionein KW - Rice SP - 154 EP - 9 JF - International journal of biological macromolecules JO - Int J Biol Macromol VL - 73 N2 - Metallothioneins (MTs) are low-molecular weight proteins with high Cys content and a high affinity for metals. Plant MTs are classified into four types based on the arrangement of Cys in their amino acid sequences. In the present study, the gene encoding OsMTI-3a, a type 3 MT found in rice, was cloned into pET41a vector. The resulting construct was transformed into the Escherichia coli strain Rosetta (DE3). Following the induction with isopropyl β-D-1-thiogalactopyranoside, the OsMTI-3a was expressed as glutathione-S-transferase (GST)-tagged fusion protein. In comparison to control strain, the cells expressing GST-OsMTI-3a accumulated more Cd(2+), Ni(2+) and Zn(2+) when they were grown in the medium containing CdCl2, NiCl2 or ZnSO4. The recombinant GST-OsMTI-3a was purified using affinity chromatography. The UV absorption spectra recorded after the reconstitution of the apo-protein with different metals confirmed that GST-OsMTI-3a was able to form complexes with Cd(2+), Ni(2+), and Zn(2+). The reaction of the protein-metal complexes with 5-5-dithiobis (2-nitrobenzoic) revealed that the order of affinity of GST-OsMTI-3a toward different metals was Ni(2+)≥Cd(2+)>Zn(2+)>Cu(2+). SN - 1879-0003 UR - https://www.unboundmedicine.com/medline/citation/25449122/Functional_characterization_of_a_type_3_metallolthionein_isoform__OsMTI_3a__from_rice_ DB - PRIME DP - Unbound Medicine ER -