Consensus model of a cyanobacterial light-dependent protochlorophyllide oxidoreductase in its pigment-free apo-form and photoactive ternary complex

Consensus model of a cyanobacterial light-dependent protochlorophyllide oxidoreductase in its pigment-free apo-form and photoactive ternary complex
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蓝藻光依赖性原叶绿素氧化还原酶的无色素脱辅基形式和光活性三元复合物的共识模型

DOI:
10.1038/s42003-019-0590-4
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发表时间:
2019
影响因子:
5.9
通讯作者:
U. Krauss
U. Krauss
中科院分区:
生物学2区
文献类型:
--
作者:
J. Schneidewind;F. Krause;M. Bocola;A.M. Stadler;K.E. Jaeger KE;U. Krauss

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光合生物使用两种不同的酶来还原原叶绿素(Pchlide)的C17 = C18双键,产生叶绿素前体叶绿素。第一,固氮酶样,光非依赖性(暗操作)Pchlide氧化还原酶,其次,光依赖性Pchlide氧化还原酶(LPOR)。对于后一种酶,尽管进行了数十年的研究,但没有结构信息。在这里,我们使用蛋白质结构建模,分子动力学(MD)模拟结合多波长分析超离心(MWA-AUC)和小角X射线散射(SAXS)实验,推导出一个共识模型的LPOR脱辅基蛋白和底物/辅因子/LPOR三元复合物。MWA-AUC和SAXS实验独立地表明,脱辅基蛋白是单体的,而三元复合物的形成诱导二聚化。SAXS指导的建模研究提供了一个全长模型的脱辅基蛋白,并提出了一个暂定模式的LPOR三元复合物的二聚化,支持发表的交联约束。我们的研究提供了LPOR结构组织的第一印象。
Photosynthetic organisms employ two different enzymes for the reduction of the C17 = C18 double bond of protochlorophyllide (Pchlide), yielding the chlorophyll precursor chlorophyllide. First, a nitrogenase-like, light-independent (dark-operative) Pchlide oxidoreductase and secondly, a light-dependent Pchlide oxidoreductase (LPOR). For the latter enzyme, despite decades of research, no structural information is available. Here, we use protein structure modelling, molecular dynamics (MD) simulations combined with multi-wavelength analytical ultracentrifugation (MWA-AUC) and small angle X-ray scattering (SAXS) experiments to derive a consensus model of the LPOR apoprotein and the substrate/cofactor/LPOR ternary complex. MWA-AUC and SAXS experiments independently demonstrate that the apoprotein is monomeric, while ternary complex formation induces dimerization. SAXS-guided modelling studies provide a full-length model of the apoprotein and suggest a tentative mode of dimerization for the LPOR ternary complex, supported by published cross-link constraints. Our study provides a first impression of the LPOR structural organization.
丙氨酸扫描诱变揭示了蛋白质表面电荷在豌豆 NADPH 催化活性和叶绿体膜缔合中的作用:原叶绿素氧化还原酶 (POR)
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发表时间: 2004
影响因子: 5.1
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期刊: BIOCHEMISTRY
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DOI: 10.1002/anie.201409881
发表时间: 2015-01-26
期刊: Angewandte Chemie (International ed. in English)
影响因子: --
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