NADPH依赖性同化亚硫酸还原酶还原酶亚基中的结构域交叉。

IF 3 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Nidhi Walia , Daniel T. Murray , Yashika Garg , Huan He , Kevin L. Weiss , Gergely Nagy , M. Elizabeth Stroupe
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引用次数: 0

摘要

来自大肠杆菌的NADPH依赖性同化亚硫酸盐还原酶(SiR)将亚硫酸盐进行六电子还原为生物可利用的硫化物。SiR由黄素蛋白(SiRFP)还原酶亚基和血蛋白(SiRHP)氧化酶亚基组成。目前还没有已知的SiR或SiRFP的高分辨率结构,因此我们还不完全了解亚基是如何相互作用来进行化学反应的。在这里,我们使用小角度中子散射来理解构象限制高流动性SiRFP八聚体为接受电子(闭合)或给予电子(开放)构象的影响,表明即使有这些构象限制,SiR仍然保持活性、柔性和不对称。根据这些散射数据,我们对SiRFP的第一个解结构进行了建模。此外,对负责SiRFP低聚的N末端52个氨基酸的计算建模表明,八个螺旋束将SiRFP亚基连接在一起形成SiR核心。最后,对封闭的SiRFP变体的质谱分析表明,SiRFP能够进行分子间结构域交叉,其中来自一种多肽的给电子结构域能够直接与另一多肽的接受电子结构域相互作用。这种结构表征表明,SiR通过SiRFP结构域之间的分子间和分子内途径进行高容量电子转移,从而从还原酶向氧化酶亚基的顺式或反式转移。这种高度冗余的电子转移潜力使该系统成为设计合成酶的潜在目标。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Domain crossover in the reductase subunit of NADPH-dependent assimilatory sulfite reductase

Domain crossover in the reductase subunit of NADPH-dependent assimilatory sulfite reductase

NADPH-dependent assimilatory sulfite reductase (SiR) from Escherichia coli performs a six-electron reduction of sulfite to the bioavailable sulfide. SiR is composed of a flavoprotein (SiRFP) reductase subunit and a hemoprotein (SiRHP) oxidase subunit. There is no known high-resolution structure of SiR or SiRFP, thus we do not yet fully understand how the subunits interact to perform their chemistry. Here, we used small-angle neutron scattering to understand the impact of conformationally restricting the highly mobile SiRFP octamer into an electron accepting (closed) or electron donating (open) conformation, showing that SiR remains active, flexible, and asymmetric even with these conformational restrictions. From these scattering data, we model the first solution structure of SiRFP. Further, computational modeling of the N-terminal 52 amino acids that are responsible for SiRFP oligomerization suggests an eight-helical bundle tethers together the SiRFP subunits to form the SiR core. Finally, mass spectrometry analysis of the closed SiRFP variant show that SiRFP is capable of inter-molecular domain crossover, in which the electron donating domain from one polypeptide is able to interact directly with the electron accepting domain of another polypeptide. This structural characterization suggests that SiR performs its high-volume electron transfer through both inter- and intramolecular pathways between SiRFP domains and, thus, cis or trans transfer from reductase to oxidase subunits. Such highly redundant potential for electron transfer makes this system a potential target for designing synthetic enzymes.

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来源期刊
Journal of structural biology
Journal of structural biology 生物-生化与分子生物学
CiteScore
6.30
自引率
3.30%
发文量
88
审稿时长
65 days
期刊介绍: Journal of Structural Biology (JSB) has an open access mirror journal, the Journal of Structural Biology: X (JSBX), sharing the same aims and scope, editorial team, submission system and rigorous peer review. Since both journals share the same editorial system, you may submit your manuscript via either journal homepage. You will be prompted during submission (and revision) to choose in which to publish your article. The editors and reviewers are not aware of the choice you made until the article has been published online. JSB and JSBX publish papers dealing with the structural analysis of living material at every level of organization by all methods that lead to an understanding of biological function in terms of molecular and supermolecular structure. Techniques covered include: • Light microscopy including confocal microscopy • All types of electron microscopy • X-ray diffraction • Nuclear magnetic resonance • Scanning force microscopy, scanning probe microscopy, and tunneling microscopy • Digital image processing • Computational insights into structure
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