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

Abstract

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.

Keywords: C-terminal Cys-rich region; Metal-thiolate cluster; Metallothionein; OsMTI-1b; Rice.