Functional analysis of metallothionein MTT5 from Tetrahymena thermophila

Journal of Cellular Biochemistry
Huanxin ZhouWei Wang

Abstract

Metallothioneins (MTs) constitute a superfamily of cysteine-rich proteins that bind heavy-metal ions by metal-thiolate clusters. Five MT genes from Tetrahymena thermophila was subdivided into 7a (MTT1, MTT3, and MTT5) and 7b (MTT2 and MTT4) subfamilies. In the study, MTT5 was knocked out in Tetrahymena. The mutant cells were sensitive to Cd2+ and Pb2+ but poorly sensitive to Cu+ . In the MTT5 knockout cells, the expression levels of MTT1 and MTT3 were significantly up-regulated under Cd2+ and Pb2+ stresses, whereas the expression levels of MTT2 and MTT4 were significantly up-regulated under Cu+ stress relative to those in the wild-type cells. Furthermore, recombinant GST-MTT5 was expressed in Escherichia coli/pGEX-MTT5 and purified by affinity chromatography. Fluorescence quenching analysis showed that apoMTT5 can bind 8 Cd2+ , 8 Pb2+ , and 12 Cu+ . The metal-binding ability of the MTT5 complex followed the order of Pb2+  > Cd2+  > Cu+ . Meanwhile, the half-maximal inhibitory concentrations of the heavy-metal ions for E. coli/pGEX-MTT5 were as follows: Cu+ (483.9 µM) > Pb2+ (410.7 µM) > Cd2+ (130.8 µM). The accumulation of Cd2+ , Pb2+ , and Cu+ in the E. coli/pGEX-MTT5 was enhanced relative to that of E. coli/pGEX-4T. Results s...Continue Reading

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Citations

Feb 6, 2018·Bioscience, Biotechnology, and Biochemistry·Huanxin ZhouWei Wang
Nov 14, 2018·Critical Reviews in Biotechnology·María E ElgueroAlejandro D Nusblat
Feb 3, 2021·Environmental Science and Pollution Research International·Anindita MitraDharmendra K Gupta

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