The conformational bases for the two functionalities of 2-cysteine peroxiredoxins as peroxidase and chaperone.
The conformational bases for the two functionalities of 2-cysteine peroxiredoxins as peroxidase and chaperone.
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DOI：
10.1093/jxb/ert184
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发表时间：
2013-08
影响因子：
6.9
通讯作者：
Dietz KJ
中科院分区：
文献类型：
--
作者：
König J;Galliardt H;Jütte P;Schäper S;Dittmann L;Dietz KJ
2-半胱氨酸过氧化物酶（2-CysPrxs）是普遍存在的并且高度丰富的蛋白质，其作为过氧化物酶、分子伴侣和硫醇氧化酶以及在氧化还原依赖性细胞信号传导中发挥多种功能。叶绿体蛋白在幼苗发育和保护光合机构中起作用。本研究旨在明确连接构象和功能。为此，工程化一组非标记的定点诱变的At 2-CysPrx变体，其模拟构象状态及其特定功能：过氧化形式（C54 D），还原形式（C54 S，C176 S），氧化形式（C54 DC 176 K），磷酸化形式（T92 D），通过干扰二聚体-二聚体界面（F84 R）和C-末端截短形式[ΔC（-20 aa）]降低寡聚化能力。这些变体完全或部分固定在其四级结构和功能，分别，并分析了它们的构象状态和过氧化物酶和伴侣活性，以及它们对过氧化的敏感性。His 6标签的存在强烈影响蛋白质的性质。ΔC变异体变得对过氧化不敏感，而T92 D和F84 R变得更敏感。C54 D变体显示最高的伴侣蛋白活性。观察到F84 R和ΔC变体的过氧化物酶活性最高。与NADP依赖性硫氧还蛋白还原酶C的有效相互作用依赖于Cys残基和C-末端尾的存在。结果表明，结构的灵活性是重要的过氧化物酶和伴侣功能之间的开关和进化保守的功能开关，而不是最大限度地提高一个单一的功能。这些变体是未来体内和体外构象特异性研究的理想工具。
2-Cysteine peroxiredoxins (2-CysPrxs) are ubiquitous and highly abundant proteins that serve multiple functions as peroxidases, chaperones, and thiol oxidases and in redox-dependent cell signalling. The chloroplast protein plays a role in seedling development and protection of the photosynthetic apparatus. This study aimed to unequivocally link conformation and function. To this end, a set of non-tagged site-directed mutagenized At2-CysPrx variants was engineered, which mimicked the conformational states and their specific functions: hyperoxidized form (C54D), reduced form (C54S, C176S), oxidized form (C54DC176K), phosphorylated form (T92D), reduced ability for oligomerization by interfering with the dimer–dimer interface (F84R) and a C-terminally truncated form [ΔC (–20 aa)]. These variants were fully or partly fixed in their quaternary structure and function, respectively, and were analysed for their conformational state and peroxidase and chaperone activity, as well as for their sensitivity to hyperoxidation. The presence of a His6-tag strongly influenced the properties of the protein. The ΔC variant became insensitive to hyperoxidation, while T92D and F84R became more sensitive. The C54D variant revealed the highest chaperone activity. The highest peroxidase activity was observed for the F84R and ΔC variants. Efficient interaction with NADP-dependent thioredoxin reductase C depended on the presence of Cys residues and the C-terminal tail. The results suggest that the structural flexibility is important for the switch between peroxidase and chaperone function and that evolution has conserved the functional switch instead of maximizing a single function. These variants are ideal tools for future conformation-specific studies in vivo and in vitro.
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影响因子：
3.8
作者：
An, Byung Chull;Lee, Seung Sik;Chung, Byung Yeoup
通讯作者：
Chung, Byung Yeoup
DOI：
10.1074/jbc.m111.298810
发表时间：
2011-12-09
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Cao Z;Tavender TJ;Roszak AW;Cogdell RJ;Bulleid NJ
通讯作者：
Bulleid NJ
影响因子：
64.8
作者：
Edgar, Rachel S.;Green, Edward W.;Zhao, Yuwei;van Ooijen, Gerben;Olmedo, Maria;Qin, Ximing;Xu, Yao;Pan, Min;Valekunja, Utham K.;Feeney, Kevin A.;Maywood, Elizabeth S.;Hastings, Michael H.;Baliga, Nitin S.;Merrow, Martha;Millar, Andrew J.;Johnson, Carl H.;Kyriacou, Charalambos P.;O'Neill, John S.;Reddy, Akhilesh B.
通讯作者：
Reddy, Akhilesh B.
影响因子：
4.8
作者：
Matsumura, Tomohiro;Okamoto, Ken;Abe, Yasuko
通讯作者：
Abe, Yasuko
影响因子：
11.5
作者：
Kim, Joo-Heon;Bogner, Paul N.;Park, Young-Mee
通讯作者：
Park, Young-Mee