Involvement of C-Terminal Histidines in Soybean PM1 Protein Oligomerization and Cu2+ Binding
Involvement of C-Terminal Histidines in Soybean PM1 Protein Oligomerization and Cu2+ Binding
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C 末端组氨酸参与大豆 PM1 蛋白寡聚化和 Cu2 结合
DOI：
10.1093/pcp/pcx046
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发表时间：
2017-06
影响因子：
4.9
通讯作者：
Zheng Yizhi
中科院分区：
文献类型：
--
作者：
Liu Guobao;Liu Ke;Gao Yang;Zheng Yizhi
晚期胚胎发生丰富（LEA）蛋白广泛分布于植物物种中，它们有助于非生物胁迫耐受性。根据保守的序列基序，LEA 蛋白可分为七组。来自大豆的 PM1 蛋白属于 Pfam LEA_1 组，此前已被证明至少部分天然未折叠，可以结合金属离子，并有可能稳定蛋白质和膜。在这里，我们研究了 PM1 C 末端结构域的作用，特别是这半蛋白质中的多个组氨酸残基。我们构建了表达全长 PM1 和两种截短形式 PM1-N 和 PM1-C 的重组质粒，它们分别代表蛋白质的 N 端和 C 端。免疫印迹和交联实验表明，全长 PM1 在体外和体内形成寡聚物和高分子量 (HMW) 复合物，而 PM1-C（而非 PM1-N）也在体外形成寡聚物和 HMW 复合物。当 PM1 和 PM1-C 中的组氨酸残基被化学修饰时，寡聚化被废除，表明组氨酸在此过程中发挥着关键作用。此外，我们证明高 Cu2+ 浓度会促进寡聚化并诱导 PM1 和 PM1-C 形成 HMW 复合物。因此，我们推测 PM1 蛋白不仅维持细胞质中的离子稳态，而且还可能在非生物胁迫期间通过在生物靶标周围形成大型寡聚分子屏蔽来稳定和保护其他蛋白质。
Late embryogenesis abundant (LEA) proteins are widely distributed among plant species, where they contribute to abiotic stress tolerance. LEA proteins can be classified into seven groups according to conserved sequence motifs. The PM1 protein from soybean, which belongs to the Pfam LEA_1 group, has been shown previously to be at least partially natively unfolded, to bind metal ions and potentially to stabilize proteins and membranes. Here, we investigated the role of the PM1 C-terminal domain and in particular the multiple histidine residues in this half of the protein. We constructed recombinant plasmids expressing full-length PM1 and two truncated forms, PM1-N and PM1-C, which represent the N- and C-terminal halves of the protein, respectively. Immunoblotting and cross-linking experiments showed that full-length PM1 forms oligomers and high molecular weight (HMW) complexes in vitro and in vivo, while PM1-C, but not PM1-N, also formed oligomers and HMW complexes in vitro. When the histidine residues in PM1 and PM1-C were chemically modified, oligomerization was abolished, suggesting that histidines play a key role in this process. Furthermore, we demonstrated that high Cu2+ concentrations promote oligomerization and induce PM1 and PM1-C to form HMW complexes. Therefore, we speculate that PM1 proteins not only maintain ion homeostasis in the cytoplasm, but also potentially stabilize and protect other proteins during abiotic stress by forming a large, oligomeric molecular shield around biological targets.
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DOI：
10.1016/j.bbamem.2012.11.031
发表时间：
2013-03
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Luna N Rahman;Fraser McKay;M. Giuliani;A. Quirk;B. Moffatt;G. Harauz;J. Dutcher
通讯作者：
Luna N Rahman;Fraser McKay;M. Giuliani;A. Quirk;B. Moffatt;G. Harauz;J. Dutcher
Pfam：氏族、网络工具和服务。
影响因子：
14.9
作者：
Finn, Robert D.;Mistry, Jaina;Schuster-Bockler, Benjamin;Griffiths-Jones, Sam;Hollich, Volker;Lassmann, Timo;Moxon, Simon;Marshall, Mhairi;Khanna, Ajay;Durbin, Richard;Eddy, Sean R.;Sonnhammer, Erik L. L.;Bateman, Alex
通讯作者：
Bateman, Alex
影响因子：
5.3
作者：
Wang M;Li P;Li C;Pan Y;Jiang X;Zhu D;Zhao Q;Yu J
通讯作者：
Yu J
影响因子：
4.9
作者：
Liu, Guobao;Xu, Hong;Zheng, Yizhi
通讯作者：
Zheng, Yizhi
影响因子：
6.9
作者：
Hara, M;Fujinaga, M;Kuboi, T
通讯作者：
Kuboi, T