Site-specific mutagenesis of yeast 2-Cys peroxiredoxin improves heat or oxidative stress tolerance by enhancing its chaperone or peroxidase function
- Authors
- Hong, Sung Hyun; Lee, Seung Sik; Chung, Jeong Min; Jung, Hyun Suk; Singh, Sudhir; Mondal, Suvendu; Jang, Ho Hee; Cho, Jae-Young; Bae, Hyeun-Jong; Chung, Byung Yeoup
- Issue Date
- Jan-2017
- Publisher
- SPRINGER WIEN
- Keywords
- 2-Cys peroxiredoxin; Heat tolerance; Molecular chaperone; Oxidative stress; Peroxidase; Site-directed mutagenesis
- Citation
- PROTOPLASMA, v.254, no.1, pp.327 - 334
- Journal Title
- PROTOPLASMA
- Volume
- 254
- Number
- 1
- Start Page
- 327
- End Page
- 334
- URI
- https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/6551
- DOI
- 10.1007/s00709-016-0948-0
- ISSN
- 0033-183X
- Abstract
- Yeast peroxiredoxin II (yPrxII) is an antioxidant enzyme that plays a protective role against the damage caused by reactive oxygen species (ROS) in Saccharomyces cerevisiae. This enzyme consists of 196 amino acids containing 2-Cys Prx with highly conserved two active cysteine residues at positions 48 and 171. The yPrxII has dual enzymatic functions as a peroxidase and molecular chaperone. To understand the effect of additional cysteine residues on dual functions of yPrxII, S79C-yPrxII and S109C-yPrxII, the substitution of Ser with Cys residue at 79 and 109 positions, respectively, was generated. S109C-yPrxII and S79C-yPrxII showed 3.7- and 2.7-fold higher chaperone and peroxidase activity, respectively, than the wild type (WT). The improvement in enzyme activity was found to be closely associated with structural changes in proteins. S109C-yPrxII had increased beta-sheet in its secondary structure and formed high-molecular-weight (HMW) as well as low-molecular-weight (LMW) complexes, but S79C-yPrxII formed only LMW complexes. HMW complexes predominantly exhibited a chaperone function, and LMW complexes showed a peroxidase function. In addition, transgenic yeast cells over-expressing Cys-substituted yPrxII showed greater tolerance against heat and oxidative stress compared to WT-yPrxII.
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