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Heterologous expression and metal-binding characterization of a type 1 metallothionein isoform (OsMTI-1b) from rice (Oryza sativa).
Protein J. 2013 Feb; 32(2):131-7.PJ

Abstract

Metallothioneins (MTs) are ubiquitous, low molecular mass and cysteine-rich proteins that play important roles in maintaining intracellular metal homeostasis, eliminating metal toxification and protecting the cells against oxidative damages. MTs are able to bind metal ions through the thiol groups of their cysteine residues. Plants have several MT isoforms which are classified into four types based on the arrangement of cysteine residues. In the present study, a rice (Oryza sativa) gene encoding type 1 MT isoform, OsMTI-1b, was inserted in vector pET41a and overexpressed in Escherichia coli as carboxy-terminal extensions of glutathione-S-transferase (GST). The recombinant protein GST-OsMTI-1b was purified using affinity chromatography and its ability to bind with Ni(2+), Cd(2+), Zn(2+) and Cu(2+) ions was analyzed. The results demonstrated that this isoform has ability to bind Ni(2+), Cd(2+) and Zn(2+) ions in vitro, whereas it has no substantial ability to bind Cu(2+) ions. From competitive reaction with 5,5'-dithiobis(2-nitrobenzoic acid), DTNB, the affinity of metal ions for recombinant form of GST-OsMTI-1b was as follows: Ni(2+)/Cd(2+) > Zn(2+) > Cu(2+).

Authors+Show Affiliations

Department of Agriculture Biotechnology, College of Agriculture, Isfahan University of Technology, 84156-83111 Isfahan, Iran.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

23385446

Citation

Nezhad, Rezvan Mohammadi, et al. "Heterologous Expression and Metal-binding Characterization of a Type 1 Metallothionein Isoform (OsMTI-1b) From Rice (Oryza Sativa)." The Protein Journal, vol. 32, no. 2, 2013, pp. 131-7.
Nezhad RM, Shahpiri A, Mirlohi A. Heterologous expression and metal-binding characterization of a type 1 metallothionein isoform (OsMTI-1b) from rice (Oryza sativa). Protein J. 2013;32(2):131-7.
Nezhad, R. M., Shahpiri, A., & Mirlohi, A. (2013). Heterologous expression and metal-binding characterization of a type 1 metallothionein isoform (OsMTI-1b) from rice (Oryza sativa). The Protein Journal, 32(2), 131-7. https://doi.org/10.1007/s10930-013-9469-2
Nezhad RM, Shahpiri A, Mirlohi A. Heterologous Expression and Metal-binding Characterization of a Type 1 Metallothionein Isoform (OsMTI-1b) From Rice (Oryza Sativa). Protein J. 2013;32(2):131-7. PubMed PMID: 23385446.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Heterologous expression and metal-binding characterization of a type 1 metallothionein isoform (OsMTI-1b) from rice (Oryza sativa). AU - Nezhad,Rezvan Mohammadi, AU - Shahpiri,Azar, AU - Mirlohi,Aghafakhr, PY - 2013/2/7/entrez PY - 2013/2/7/pubmed PY - 2013/8/2/medline SP - 131 EP - 7 JF - The protein journal JO - Protein J VL - 32 IS - 2 N2 - Metallothioneins (MTs) are ubiquitous, low molecular mass and cysteine-rich proteins that play important roles in maintaining intracellular metal homeostasis, eliminating metal toxification and protecting the cells against oxidative damages. MTs are able to bind metal ions through the thiol groups of their cysteine residues. Plants have several MT isoforms which are classified into four types based on the arrangement of cysteine residues. In the present study, a rice (Oryza sativa) gene encoding type 1 MT isoform, OsMTI-1b, was inserted in vector pET41a and overexpressed in Escherichia coli as carboxy-terminal extensions of glutathione-S-transferase (GST). The recombinant protein GST-OsMTI-1b was purified using affinity chromatography and its ability to bind with Ni(2+), Cd(2+), Zn(2+) and Cu(2+) ions was analyzed. The results demonstrated that this isoform has ability to bind Ni(2+), Cd(2+) and Zn(2+) ions in vitro, whereas it has no substantial ability to bind Cu(2+) ions. From competitive reaction with 5,5'-dithiobis(2-nitrobenzoic acid), DTNB, the affinity of metal ions for recombinant form of GST-OsMTI-1b was as follows: Ni(2+)/Cd(2+) > Zn(2+) > Cu(2+). SN - 1875-8355 UR - https://www.unboundmedicine.com/medline/citation/23385446/Heterologous_expression_and_metal_binding_characterization_of_a_type_1_metallothionein_isoform__OsMTI_1b__from_rice__Oryza_sativa__ L2 - https://link.springer.com/article/10.1007/s10930-013-9469-2 DB - PRIME DP - Unbound Medicine ER -