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首页> 外文期刊>Biochemistry >Autophosphorylation of Arabidopsis Nucleoside Diphosphate Kinase 2 Occurs Only on Its Active Histidine Residue
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Autophosphorylation of Arabidopsis Nucleoside Diphosphate Kinase 2 Occurs Only on Its Active Histidine Residue

机译：拟南芥核苷二磷酸三磷酸激酶2仅发生在其活性组氨酸残基上发生自磷酸化

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摘要

Arabidopsis nucleoside diphosphate kinase 2 (NDPK2) is a component in the phytochrome-mediated light signaling.In the present study,its autophosphorylation was investigated.Acid-stable and alkali-stable phosphorylated residues were analyzed under two different conditions.Results revealed that NDPK2 is phosphorylated only on its active histidine residue His197 and the presence of serine/threonine phosphorylation is an experimental artifact due to the harsh condition applied in the treatment of the phosphorylated protein sample.To resolve the controversy of whether serine/threonine phosphorylation of NDPK occurs as has been suggested by other NDPK studies,NDPK2 putative phosphorylation site mutants were generated and examined.No serine/threonine phosphorylation was identified in NDPK2 or implicated in its enzymatic activity.Further studies indicated that the low enzymatic activity and autophosphorylation level of NDPK2 mutant S199A are shown to be due to a damaged H-bonding with the active histidine residue His197 in the nucleotide-binding pocket.In addition,NDPK2 Kpn loop mutant T182A was found to possess an extremely low enzymatic activity and almost no autophosphorylation,suggesting the importance of the oligomeric states of NDPK2 in NDPK2 functioning.
机译：拟南芥核苷激酶2（NDPK2）是植物中介导的光信号传递中的组分。在本研究中，研究其自磷酸化。在两个不同的条件下分析其自磷酸化。酸稳定，碱稳定的磷酸化残基分析。结果显示NDPK2是仅在其活性组氨酸残留物上磷酸化并且丝氨酸/苏氨酸磷酸化的存在是一种实验伪影，其由于在治疗磷酸化的蛋白样品中施加的苛刻条件而存在。解决NDPK的丝氨酸/苏氨酸磷酸化的争论已经提出了其他NDPK研究，产生并检测了NDPK2推定的磷酸化位点突变体。在NDPK2中鉴定了NDPK2的丝氨酸/苏氨酸磷酸化，或者在其酶活性中涉及。结果表明，显示了NDPK2突变体S199A的低酶活性和自磷酸化水平。显示出NDPK2突变体S199A的低酶活性和自磷酸化水平由于与Acti损坏的H键合VE组氨酸残留在核苷酸结合口袋中的197。另外，发现NDPK2 KPN环突变体T182A具有极低的酶活性，几乎没有自磷酸化，表明NDPK2在NDPK2中的低聚状态的重要性。

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  • 来源
    《Biochemistry》 |2006年第6期|共4页
  • 作者

    Yu Shen; Jeong-Il Kim; Pill-Soon Song;

  • 作者单位

    Department of Chemistry University of Nebraska-Lincoln Lincoln Nebraska 68588-0304 Environmental Biotechnology Research Center Gyeongsang National University Jinju 660-701 Korea Kumho Life and Environmental Science Laboratory 1 Oryong-Dong Gwangju 5;

    Department of Chemistry University of Nebraska-Lincoln Lincoln Nebraska 68588-0304 Environmental Biotechnology Research Center Gyeongsang National University Jinju 660-701 Korea Kumho Life and Environmental Science Laboratory 1 Oryong-Dong Gwangju 5;

    Department of Chemistry University of Nebraska-Lincoln Lincoln Nebraska 68588-0304 Environmental Biotechnology Research Center Gyeongsang National University Jinju 660-701 Korea Kumho Life and Environmental Science Laboratory 1 Oryong-Dong Gwangju 5;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物化学;
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