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Functional characterization of a type 2 metallothionein isoform (OsMTI-2b) from rice.
Int J Biol Macromol. 2016 Jul; 88:491-6.IJ

Abstract

Metallothioneins (MTs) are a family of Cys-rich, low molecular weight, cytoplasmic metal binding proteins. MTs are present in all eukaryotes as well as some prokaryotes. Plant MTs are divided into four types based on Cys distribution pattern in their amino acid sequences. In the present work, the gene encoding OsMTI-2b, a type 2 MT found in rice, was cloned into pET41a vector. The resulting construct was transformed into Escherichia coli strain Rosetta (DE3). Following the induction with Isopropyl β-d-1-thiogalactopyranoside the OsMTI-2b was expressed as carboxyl-terminal extensions of glutathione-S-transferase (GST-tag), a 6His-tag, and an S-tag. The expressed recombinant fusion protein was named GST-OsMTI-2b. As compared with control, transgenic E. coli cells expressing GST-OsMTI-2b accumulated more Pb(2+), Ni(2+), Cd(2+), Zn(2+) and Cu(2+) from culture medium and showed increased tolerance against these metals. Furthermore the E. coli cells expressing OsMTI-2b accumulated significantly higher Pb(2+) than previously made strains which expressing other rice OsMT isoforms. The recombinant GST-OsMTI-2b was purified using affinity chromatography. According to in vitro assays the protein GST-OsMTI-2b was able to form complexes with Pb(2+), Ni(2+), Cd(2+) and Zn(2+). However, the binding ability for the different metals differed in the order: Pb(2+)>Cd(2+)>Zn(2+)>Ni(2+).

Authors+Show Affiliations

Department of Biotechnology, College of Agriculture, Isfahan University of Technology, Isfahan, Iran.Department of Biotechnology, College of Agriculture, Isfahan University of Technology, Isfahan, Iran. Electronic address: a.shahpiri@cc.iut.ac.ir.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27079330

Citation

Pirzadeh, Soheil, and Azar Shahpiri. "Functional Characterization of a Type 2 Metallothionein Isoform (OsMTI-2b) From Rice." International Journal of Biological Macromolecules, vol. 88, 2016, pp. 491-6.
Pirzadeh S, Shahpiri A. Functional characterization of a type 2 metallothionein isoform (OsMTI-2b) from rice. Int J Biol Macromol. 2016;88:491-6.
Pirzadeh, S., & Shahpiri, A. (2016). Functional characterization of a type 2 metallothionein isoform (OsMTI-2b) from rice. International Journal of Biological Macromolecules, 88, 491-6. https://doi.org/10.1016/j.ijbiomac.2016.04.021
Pirzadeh S, Shahpiri A. Functional Characterization of a Type 2 Metallothionein Isoform (OsMTI-2b) From Rice. Int J Biol Macromol. 2016;88:491-6. PubMed PMID: 27079330.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Functional characterization of a type 2 metallothionein isoform (OsMTI-2b) from rice. AU - Pirzadeh,Soheil, AU - Shahpiri,Azar, Y1 - 2016/04/11/ PY - 2015/10/29/received PY - 2016/04/05/revised PY - 2016/04/07/accepted PY - 2016/4/16/entrez PY - 2016/4/16/pubmed PY - 2017/3/8/medline KW - Heavy metal tolerance KW - Metal-binding ability KW - Metallothionein KW - Rice KW - Type 2 MT SP - 491 EP - 6 JF - International journal of biological macromolecules JO - Int J Biol Macromol VL - 88 N2 - Metallothioneins (MTs) are a family of Cys-rich, low molecular weight, cytoplasmic metal binding proteins. MTs are present in all eukaryotes as well as some prokaryotes. Plant MTs are divided into four types based on Cys distribution pattern in their amino acid sequences. In the present work, the gene encoding OsMTI-2b, a type 2 MT found in rice, was cloned into pET41a vector. The resulting construct was transformed into Escherichia coli strain Rosetta (DE3). Following the induction with Isopropyl β-d-1-thiogalactopyranoside the OsMTI-2b was expressed as carboxyl-terminal extensions of glutathione-S-transferase (GST-tag), a 6His-tag, and an S-tag. The expressed recombinant fusion protein was named GST-OsMTI-2b. As compared with control, transgenic E. coli cells expressing GST-OsMTI-2b accumulated more Pb(2+), Ni(2+), Cd(2+), Zn(2+) and Cu(2+) from culture medium and showed increased tolerance against these metals. Furthermore the E. coli cells expressing OsMTI-2b accumulated significantly higher Pb(2+) than previously made strains which expressing other rice OsMT isoforms. The recombinant GST-OsMTI-2b was purified using affinity chromatography. According to in vitro assays the protein GST-OsMTI-2b was able to form complexes with Pb(2+), Ni(2+), Cd(2+) and Zn(2+). However, the binding ability for the different metals differed in the order: Pb(2+)>Cd(2+)>Zn(2+)>Ni(2+). SN - 1879-0003 UR - https://www.unboundmedicine.com/medline/citation/27079330/Functional_characterization_of_a_type_2_metallothionein_isoform__OsMTI_2b__from_rice_ DB - PRIME DP - Unbound Medicine ER -