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首页> 外文学位 >Phosphate-starvation-inducible vacuolar purple acid phosphatase from Arabidopsis thaliana suspension cell cultures.
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Phosphate-starvation-inducible vacuolar purple acid phosphatase from Arabidopsis thaliana suspension cell cultures.

机译：来自拟南芥悬浮细胞培养物中的磷酸盐饥饿诱导的液泡紫色酸性磷酸酶。

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摘要

Acid phosphatases (APs) function in the production and recycling of Pi, a crucial but limiting macronutrient for plant growth and metabolism. Quantification of immunoblots revealed that the ∼4-fold increase in AP activity that occurred following 7-d of Pi deprivation of Arabidopsis thaliana suspension cells was paralleled by a similar increase in the amount of an anti-(tomato purple AP (PAP))-IgG immunoreactive 55-kDa polypeptide. An AP from Pi deficient (-Pi) Arabidopsis cells was purified 957-fold to homogeneity and a final phosphoenolpyruvate-hydrolyzing specific activity of 421 units/mg protein. The final AP preparation was pink in colour and insensitive to tartrate inhibition, indicating that it is a PAP. BLAST analysis of its N-terminal sequence demonstrated that it is encoded by 1 of 29 putative Arabidopsis PAP genes, specifically AtPAP26, and that a 30 amino acid signal peptide is cleaved from the preprotein following its translocation into the vacuole. AtPAP26 exists as a ∼100 kDa homodimer of 55-kDa glycosylated subunits, and displayed a pH-activity optimum of 5.6, activation by Mg2+, but potent inhibition by Zn2+, Fe2+, molybdate, and phosphate. Furthermore, the AP exhibited significant peroxidase activity. Immunoblotting of roots and shoots from Pi sufficient (+Pi) vs. -Pi Arabidopsis seedlings indicated that this PAP may be root specific under short-term Pi starvation. This is the first Pi starvation inducible intracellular Arabidopsis PAP that has been purified and characterized. It is hypothesized that this PAP plays a pivotal role in Pi remobilization and scavenging from intracellular P-monoesters in -Pi Arabidopsis.
机译：酸性磷酸酶（AP）在Pi的生产和循环中发挥作用，Pi是植物生长和代谢的关键但有限的大量营养素。免疫印迹的定量分析表明，在拟南芥悬浮细胞的Pi剥夺7天后，AP活性增加了约4倍，同时抗-（番茄紫色AP（PAP））- IgG免疫反应性55-kDa多肽。将来自Pi缺陷型（-Pi）拟南芥细胞的AP纯化957倍至均质，最终水解磷酸烯醇丙酮酸的比活性为421单位/ mg蛋白质。最终的AP制剂为粉红色，对酒石酸抑制不敏感，表明它是PAP。 BLAST对其N-末端序列的分析表明，它由29个推测的拟南芥PAP基因中的1个，特别是AtPAP26编码，并且在其转移到液泡中后，从前蛋白上切割了30个氨基酸的信号肽。 AtPAP26以55 kDa糖基化亚基的〜100 kDa同型二聚体存在，最适pH值为5.6，被Mg2 +激活，但被Zn2 +，Fe2 +，钼酸盐和磷酸盐有效抑制。此外，AP显示出显着的过氧化物酶活性。足够的Pi（-Pi）与-Pi拟南芥幼苗的根和芽的免疫印迹表明，该PAP在短期Pi饥饿下可能是根特异性的。这是已经纯化和鉴定的第一个Pi饥饿诱导的细胞内拟南芥PAP。据推测，该PAP在Pi的转运和清除-Pi拟南芥中的细胞内P-单酯中起关键作用。

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  • 作者

    Veljanovski, Vasko.;

  • 作者单位

    Queen's University at Kingston (Canada).;

  • 授予单位 Queen's University at Kingston (Canada).;
  • 学科 Biology Plant Physiology.
  • 学位 M.Sc.
  • 年度 2005
  • 页码 80 p.
  • 总页数 80
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 植物学;
  • 关键词

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