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Metalation of a rice type 1 metallothionein isoform (OsMTI-1b).
Protein Expr Purif. 2020 11; 175:105719.PE

Abstract

The simultaneously functions of Metallothioneins (MTs) are relied on their metalation mechanisms that can be divided into non-cooperative, weakly cooperative and strongly cooperative mechanisms. In this study, we recombinantly synthesized OsMTI-1b, N- and C-terminal Cys-rich regions as glutathione-S-transferase (GST)-fusion proteins in E. coli. In comparison with control strains (The E. coli cells containing pET41a without gene), transgenic E. coli cells showed more tolerance against Cd2+ and Zn2+. The recombinant GST-proteins were purified using affinity chromatography. According to in vitro assays, the recombinant proteins showed a higher binding ability to Cd2+ and Zn2+. However, the affinity of apo-proteins to Cu2+ ions were very low. The coordination of Cd2+ ions in OsMTI-1b demonstrates a strongly cooperative mechanism with a priority for the C-terminal Cys-rich region that indicates the detoxifying of heavy metals as main role of P1 subfamily of MTs. While the metalation with Zn2+ conformed to a weakly cooperative mechanism with a specificity to N-terminal Cys-rich region. It implies the specific function of OsMTI-1b is involved in zinc homeostasis. Nevertheless, a non-cooperative metalation mechanism was perceived for Cu2+ that suggests the fully metalation does not occur and OsMTI-1b cannot play a significant role in dealing with Cu2+ ions.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Malekzadeh R
Department of Medical Biotechnology, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, 88157-13471, Iran; Student Research Committee, Shahrekord University of Medical Sciences, Shahrekord, Iran. Electronic address: st-malekzadeh.r@skums.ac.ir.
Shahpiri A
Department of Biotechnology, College of Agriculture, Isfahan University of Technology, Isfahan, 84156-83111, Iran.
Siapoush S
Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Science, Tabriz, 51664-15731, Iran.

MeSH

CadmiumChromatography, AffinityCopperEscherichia coliMetallothioneinOryzaPlant ProteinsRecombinant Fusion ProteinsZinc

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32750405

Citation

Malekzadeh, Rahim, et al. "Metalation of a Rice Type 1 Metallothionein Isoform (OsMTI-1b)." Protein Expression and Purification, vol. 175, 2020, p. 105719.
Malekzadeh R, Shahpiri A, Siapoush S. Metalation of a rice type 1 metallothionein isoform (OsMTI-1b). Protein Expr Purif. 2020;175:105719.
Malekzadeh, R., Shahpiri, A., & Siapoush, S. (2020). Metalation of a rice type 1 metallothionein isoform (OsMTI-1b). Protein Expression and Purification, 175, 105719. https://doi.org/10.1016/j.pep.2020.105719
Malekzadeh R, Shahpiri A, Siapoush S. Metalation of a Rice Type 1 Metallothionein Isoform (OsMTI-1b). Protein Expr Purif. 2020;175:105719. PubMed PMID: 32750405.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Metalation of a rice type 1 metallothionein isoform (OsMTI-1b). AU - Malekzadeh,Rahim, AU - Shahpiri,Azar, AU - Siapoush,Samaneh, Y1 - 2020/08/01/ PY - 2020/04/18/received PY - 2020/06/11/revised PY - 2020/07/25/accepted PY - 2020/8/5/pubmed PY - 2021/1/29/medline PY - 2020/8/5/entrez KW - Cys-rich region KW - Heavy metal KW - Metalation mechanism KW - Protein-metal interaction SP - 105719 EP - 105719 JF - Protein expression and purification JO - Protein Expr Purif VL - 175 N2 - The simultaneously functions of Metallothioneins (MTs) are relied on their metalation mechanisms that can be divided into non-cooperative, weakly cooperative and strongly cooperative mechanisms. In this study, we recombinantly synthesized OsMTI-1b, N- and C-terminal Cys-rich regions as glutathione-S-transferase (GST)-fusion proteins in E. coli. In comparison with control strains (The E. coli cells containing pET41a without gene), transgenic E. coli cells showed more tolerance against Cd2+ and Zn2+. The recombinant GST-proteins were purified using affinity chromatography. According to in vitro assays, the recombinant proteins showed a higher binding ability to Cd2+ and Zn2+. However, the affinity of apo-proteins to Cu2+ ions were very low. The coordination of Cd2+ ions in OsMTI-1b demonstrates a strongly cooperative mechanism with a priority for the C-terminal Cys-rich region that indicates the detoxifying of heavy metals as main role of P1 subfamily of MTs. While the metalation with Zn2+ conformed to a weakly cooperative mechanism with a specificity to N-terminal Cys-rich region. It implies the specific function of OsMTI-1b is involved in zinc homeostasis. Nevertheless, a non-cooperative metalation mechanism was perceived for Cu2+ that suggests the fully metalation does not occur and OsMTI-1b cannot play a significant role in dealing with Cu2+ ions. SN - 1096-0279 UR - https://www.unboundmedicine.com/medline/citation/32750405/Metalation_of_a_rice_type_1_metallothionein_isoform__OsMTI_1b__ DB - PRIME DP - Unbound Medicine ER -