Independent metal-thiolate cluster formation in C-terminal Cys-rich region of a rice type 1 metallothionein isoform

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• 作者：Rahim Malekzadeh ; Azar Shahpiri ; ^{a.shahpiri@cc.iut.ac.ir}
• 关键词：MT ; metallothionein ; Os ; Oryza sativa ; IPTG ; isopropyl β-d-1-thiogalactopyranoside ; GST ; glutathione-s-transferase ; LB ; Luria-Bertani ; DTNB ; 5 ; 50-dithiobis(2nitrobenzoic) ; TNB ; 2-nitro-5-thiobenzoic acid
• 刊名：International Journal of Biological Macromolecules
• 出版年：2017
• 出版时间：March 2017
• 年：2017
• 卷：96
• 期：Complete
• 页码：436-441
• 全文大小：1364 K
• 卷排序：96

文摘

In this study we examined the independent self assembly of metal-binding in C-terminal Cys- rich region of a type 1 metallothionein (MT) isoform from rice (OsMTI-1b). To this end the N-terminal of OsMTI-1b (C-OsMTI-1b) was heterologously expressed in Escherichia coli as fusion protein with glutathione-S-transferase (GST). As compared with control (The E. coli cells containing pET41a without gene), transgenic E. coli cells expressing GST-C-OsMTI-1b accumulated more Ni2+, Cd2+, and Zn2+ from culture medium and showed increased tolerance against these metals. The recombinant GST-C-OsMTI-1b was purified using affinity chromatography. According to in vitro assays the protein GST-C-OsMTI-1b was able to form complexes with Ni2+, Cd2+ and Zn2+. These results demonstrate the formation of independent metal-thiolate cluster at C-terminal Cys-rich region of OsMTI-1b without participation of N-terminal Cys-rich region.