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钾离子通过拟南芥摄取器 AUX1 介导的 IAA 运输的研究在异源酵母系统中进行。

Potassium Stimulation of IAA Transport Mediated by the Arabidopsis Importer AUX1 Investigated in a Heterologous Yeast System.

机构信息

Department of Life Science and Institute of Bioinformatics and Structural Biology, College of Life Science, National Tsing Hua University, No. 101, Sec. 2, Guangfu Rd. East Dist., Hsin Chu, 30013, Taiwan, Republic of China.

Department of Biotechnology and Bioindustry Sciences, National Cheng Kung University, Tainan, 701, Taiwan, Republic of China.

出版信息

J Membr Biol. 2019 Jun;252(2-3):183-194. doi: 10.1007/s00232-019-00065-6. Epub 2019 May 3.

Abstract

Auxin regulates diverse processes involved in plant growth and development. AUX1 is the first identified and most widely investigated auxin importer, and plays an important role in root gravitropism and the development of lateral root and root hair. However, the regulation of auxin transport by AUX1 is still not well understood. In this study, we examined the effect of metal ions on AUX1 transport function and found that the activity could be specifically stimulated four times by K. Further experiments revealed the preference of KF on the enhancement of transport activity of AUX1 over KCl, KBr, and KI. In addition, the interaction between K and AUX1 confers AUX1 more resistant to thermal stress but more vulnerable to proteolysis. Conventional chemical modification indicated that the extracellular acidic amino acids of AUX1 play a key role in the K stimulation. Site-specific mutagenesis showed that the replacement of Asp, Asp, and Asp of AUX1 to alanine deteriorated the K-stimulated auxin transport. By contrast, when these residues were mutated to glutamate, lysine, or asparagine, only the D312E variant restored the IAA transport activity to the wild-type level. It is thus convinced that D312 is presumably the most promising residue for the K stimulation on AUX1.

摘要

生长素调节植物生长和发育过程中的多种过程。AUX1 是第一个被鉴定出的也是研究最多的生长素输入载体,在根向地性和侧根及根毛发育中发挥重要作用。然而,AUX1 对生长素运输的调节仍不明确。在这项研究中,我们研究了金属离子对 AUX1 运输功能的影响,发现 K 可特异性地将 AUX1 的活性刺激四倍。进一步的实验揭示了 KF 对 AUX1 运输活性的增强作用优于 KCl、KBr 和 KI。此外,K 与 AUX1 的相互作用使 AUX1 对热应激更具抗性,但对蛋白水解更敏感。常规化学修饰表明,AUX1 的细胞外酸性氨基酸在 K 刺激中起关键作用。定点突变显示,将 AUX1 的 Asp、Asp 和 Asp 替换为丙氨酸会使 K 刺激的生长素运输恶化。相比之下,当这些残基突变为谷氨酸、赖氨酸或天冬酰胺时,只有 D312E 变体将 IAA 运输活性恢复到野生型水平。因此可以肯定的是,D312 可能是 AUX1 上 K 刺激的最有希望的残基。

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