title: Site-specific mutagenesis of yeast 2-Cys peroxiredoxin improves heat or oxidative stress tolerance by enhancing its chaperone or peroxidase function.
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 β-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.
authors: Research Division for Biotechnology, Advanced Radiation Technology Institute (ARTI), Korea Atomic Energy Research Institute (KAERI), 29 Geumgu-gil, Jeongeup, 56212, Korea.|Department of Bioenergy Science and Technology, Chonnam National University, Gwangju, 61186, Republic of Korea.
authors: Sung Hyun
authors: SH
authors: Hong
authors: Research Division for Biotechnology, Advanced Radiation Technology Institute (ARTI), Korea Atomic Energy Research Institute (KAERI), 29 Geumgu-gil, Jeongeup, 56212, Korea.
authors: Seung Sik
authors: SS
authors: Lee
authors: Department of Biochemistry, College of Natural Sciences, Kangwon National University, 1, Kangwondaehak-gil, Chuncheon-si, Gangwon-do, 24341, Korea.
authors: Jeong Min
authors: JM
authors: Chung
authors: Department of Biochemistry, College of Natural Sciences, Kangwon National University, 1, Kangwondaehak-gil, Chuncheon-si, Gangwon-do, 24341, Korea.
authors: Hyun Suk
authors: HS
authors: Jung
authors: Research Division for Biotechnology, Advanced Radiation Technology Institute (ARTI), Korea Atomic Energy Research Institute (KAERI), 29 Geumgu-gil, Jeongeup, 56212, Korea.
authors: Sudhir
authors: S
authors: Singh
authors: Research Division for Biotechnology, Advanced Radiation Technology Institute (ARTI), Korea Atomic Energy Research Institute (KAERI), 29 Geumgu-gil, Jeongeup, 56212, Korea.
authors: Suvendu
authors: S
authors: Mondal
authors: Department of Molecular Medicine, Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Incheon, 21936, Republic of Korea.
authors: Ho Hee
authors: HH
authors: Jang
authors: Department of Bioenvironmental Chemistry, Chonbuk National University, 567, Baekje-daero, Deokjin-gu, Jeonju, 54896, Korea.
authors: Jae-Young
authors: JY
authors: Cho
authors: Department of Bioenergy Science and Technology, Chonnam National University, Gwangju, 61186, Republic of Korea.
authors: Hyeun-Jong
authors: HJ
authors: Bae
authors: Research Division for Biotechnology, Advanced Radiation Technology Institute (ARTI), Korea Atomic Energy Research Institute (KAERI), 29 Geumgu-gil, Jeongeup, 56212, Korea. bychung@kaeri.re.kr.
authors: Byung Yeoup
authors: BY
authors: Chung
mesh_terms: D005816:Genetic Complementation Test; D018869:Heat-Shock Response; D057927:Hydrophobic and Hydrophilic Interactions; D018832:Molecular Chaperones / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D018384:Oxidative Stress; D009195:Peroxidase / Q000378:metabolism; D054464:Peroxiredoxins / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*; D013312:Stress, Physiological
chemical_list: D018832:Molecular Chaperones; D050505:Mutant Proteins; D029701:Saccharomyces cerevisiae Proteins; D054464:Peroxiredoxins; D009195:Peroxidase
keywords: 2-Cys peroxiredoxin; Heat tolerance; Molecular chaperone; Oxidative stress; Peroxidase; Site-directed mutagenesis