Insight into the oligomeric structure of PORA from A. thaliana
Insight into the oligomeric structure of PORA from A. thaliana
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DOI：
10.1016/j.bbapap.2016.09.015
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发表时间：
2016-12-01
影响因子：
3.2
通讯作者：
Kruk, Jerzy
中科院分区：
文献类型：
--
作者：
Gabruk, Michal;Nowakowska, Zuzanna;Kruk, Jerzy
光依赖性原叶绿内酯氧化还原酶（POR， E.C. 1.3.1.33）是一种直接需要光来进行生化反应的植物酶。在本文中，我们证实了POR在与底物结合之前在溶液中形成大的低聚物。我们使用不同的技术进行研究：交联，天然凝胶电泳和FRET测量。交联产物的质谱分析提供了关于POR低聚物组织的第一个结构数据。结果表明，邻近亚基的催化基在与底物结合后相互靠近。此外，我们发现了两个干扰其寡聚化特性的POR突变：Delta 85-88和Delta 240-270。此外，对Delta 189-194、Delta 240-270、Delta 318-331和Delta 392-393突变，催化活性完全丧失。(C) 2016 Elsevier B.V.版权所有
Light-dependent protochlorophyllide oxidoreductase (POR, E.C. 1.3.1.33) is a plant enzyme that directly needs light to conduct a biochemical reaction. In the present paper we confirmed that POR forms large oligomers in solution before binding of substrates. We carried out the research using different techniques: cross-linking, native gel electrophoresis and FRET measurements. Mass spectrometry analysis of the cross-link products provided the first structural data about the organisation of the oligomer of POR. The results indicated that the catalytic motifs of the adjacent subunits become close to each other upon binding of substrates. Moreover, we identified two mutations of POR that disturbed its oligomerisation properties: Delta 85-88 and Delta 240-270. Additionally, a complete loss of the catalytic activity was observed for the following mutations: Delta 189-194, Delta 240-270, Delta 318-331 and Delta 392-393. (C) 2016 Elsevier B.V. All rights reserved.