Ca2+ 结合使二聚体双氧化酶的截短 EF-手结构域转变为单体

IF 3.3 3区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Chin-Chuan Wei , Amena Abdul Razzak , Hadis Ghasemi , Rahil Khedri , Alexandria Fraase
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引用次数: 0

摘要

双氧化酶(Duox)产生的过氧化氢对甲状腺激素的合成至关重要。Duox 的活化涉及 Ca2+ 与其 EF 手域(EFD)的结合,EFD 包含两个 EF 手(EFs)。在这项研究中,我们利用光谱法、量热法、电泳迁移率法和凝胶过滤法对截短的 EFD 进行了表征,以获得其 Ca2+ 结合的热力学和动力学,并评估相关的构象变化。我们的研究结果表明,其第 2 EF-手(EF2)与 Ca2+ 的结合放热较强(Ka = 107 M-1),而 EF1 与 Ca2+ 的结合较弱(Ka = 105 M-1),导致其带负电荷的残基被掩埋。Ca2+ 与 EFD 的结合导致其结构稳定,熔化温度从 67°C 上升到 99°C,并诱导其结构从二聚体形式转变为单体形式。EF2似乎在其apo形式的二聚体形成中起作用,而Ca2+结合的EF1的疏水暴露对其holo形式的二聚体形成至关重要。这一结果与低温电子显微镜获得的结构一致,表明 EFD 在与 Ca2+ 结合后具有疏水斑块的稳定结构对其 Duox 结构域-结构域之间的电子传递相互作用至关重要。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Ca2+ binding shifts dimeric dual oxidase's truncated EF-hand domain to monomer

Ca2+ binding shifts dimeric dual oxidase's truncated EF-hand domain to monomer

Hydrogen peroxide, produced by Dual Oxidase (Duox), is essential for thyroid hormone synthesis. Duox activation involves Ca2+ binding to its EF-hand Domain (EFD), which contains two EF-hands (EFs). In this study, we characterized a truncated EFD using spectrometry, calorimetry, electrophoretic mobility, and gel filtration to obtain its Ca2+ binding thermodynamic and kinetics, as well as to assess the associated conformational changes. Our results revealed that its 2nd EF-hand (EF2) exhibits a strong exothermic Ca2+ binding (Ka = 107 M−1) while EF1 shows a weaker binding (Ka = 105 M−1), resulting in the burial of its negatively charged residues. The Ca2+ binding to EFD results in a stable structure with a melting temperature shifting from 67 to 99 °C and induces a structural transition from a dimeric to monomeric form. EF2 appears to play a role in dimer formation in its apo form, while the hydrophobic exposure of Ca2+-bound-EF1 is crucial for dimer formation in its holo form. The result is consistent with structures obtained from Cryo-EM, indicating that a stable structure of EFD with hydrophobic patches upon Ca2+ binding is vital for its Duox's domain-domain interaction for electron transfer.

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来源期刊
Biophysical chemistry
Biophysical chemistry 生物-生化与分子生物学
CiteScore
6.10
自引率
10.50%
发文量
121
审稿时长
20 days
期刊介绍: Biophysical Chemistry publishes original work and reviews in the areas of chemistry and physics directly impacting biological phenomena. Quantitative analysis of the properties of biological macromolecules, biologically active molecules, macromolecular assemblies and cell components in terms of kinetics, thermodynamics, spatio-temporal organization, NMR and X-ray structural biology, as well as single-molecule detection represent a major focus of the journal. Theoretical and computational treatments of biomacromolecular systems, macromolecular interactions, regulatory control and systems biology are also of interest to the journal.
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