Cloning and characterization of Thioredoxin 1 from the Cnidarian Hydra. (15th September 2021)

• Record Type: Journal Article
• Authors: Perween, Nusrat; Pekhale, Komal; Haval, Gauri; Mittal, Smriti; Ghaskadbi, Surendra; Ghaskadbi, Saroj S
• Is Part Of: Journal of biochemistry. Volume 171:Number 1(2022)
• Page Start: 41
• Page End: 51
• Publication Date: 2021-09-15
• Subjects: Evolutionary conservation -- Hydra -- redox regulation -- regeneration -- Trx1; Biochemistry -- Periodicals; Biochemistry -- Periodicals; Electronic journals; 572.05
• DOI: 10.1093/jb/mvab092
• Languages: English
• ISSNs: 0021-924X

Abstract

Abstract: Thioredoxins, small disulphide-containing redox proteins, play an important role in the regulation of cellular thiol redox balance through their disulfide reductase activity. In this study, we have identified, cloned, purified and characterized thioredoxin 1 ( Hv Trx1) from the Cnidarian Hydra vulgaris Ind-Pune. Bioinformatics analysis revealed that Hv Trx1 contains an evolutionarily conserved catalytic active site Cys-Gly-Pro-Cys and shows a closer phylogenetic relationship with vertebrate Trx1. Optimum pH and temperature for enzyme activity of purified Hv Trx1 was found to be pH 7.0 and 25°C, respectively. Enzyme activity decreased significantly at acidic or alkaline pH as well as at higher temperatures. Hv Trx1 was found to be expressed ubiquitously in whole mount in situ hybridization. Treatment of Hydra with hydrogen peroxide (H2 O2 ), a highly reactive oxidizing agent, led to a significant increase in gene expression and enzyme activity of Trx1. Further experiments using PX12, an inhibitor of Trx1, indicated that Trx1 plays an important role in regeneration in Hydra . Finally, by using growth assay in Escherichia coli and wound healing assay in human colon cancer cells, we demonstrate that Hv Trx1 is functionally active in both prokaryotic and eukaryotic heterologous systems. Graphical Abstract: