HSPA8 与血红素结合后转变为血红素过氧化物酶催化血红素聚合的作用

IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY The Protein Journal Pub Date : 2023-12-08 DOI:10.1007/s10930-023-10167-9
Alok Kumar Pandey, Vishal Trivedi
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引用次数: 0

摘要

血红素是血红蛋白降解的副产品,会对细胞造成氧化损伤。血红素积聚后,几种蛋白质会被招募来进行血红素解毒,其中血红素加氧酶发挥着关键作用。伴侣蛋白主要通过防止蛋白质降解和折叠来发挥保护作用。众所周知,它们在类似情况下也有其他次要作用。为了发现伴侣在血红素应激过程中的次要作用,我们研究了伴侣 HSPA8 在血红素解毒过程中的作用。室内研究表明,HSPA8 有一个定义明确的生物功能环境来结合 hemin。通过光差光谱法,我们发现 HSPA8 通过其 N 端结构域与 hemin 结合,其 Kd 值为 5.9 ± 0.04 µM,并转化为一种血蛋白。我们以愈创木酚为底物,测试了该血蛋白是否具有过氧化物酶活性。形成的复合物与 H2O2 发生反应,表现出典型的过氧化物酶活性,能够氧化芳香族和卤化物底物。HSPA8 通过其 N 端结构域具有剂量依赖性地催化血红素聚合。红外光谱结果显示,形成的聚合物在结构上与 β-血红素相似,表明其具有共价性质。聚合机制通过光学光谱、自旋阱和活性抑制实验进行了检验。结果表明,聚合是通过过氧化物酶-H2O2 系统进行的,其中涉及单电子转移机制,以及自由基和自由基-自由基相互作用的形成。它强调了 HSPA8-hemin 复合物在疟疾、镰状细胞病等病理情况下可能发挥的细胞保护功能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Role Transformation of HSPA8 to Heme-peroxidase After Binding Hemin to Catalyze Heme Polymerization

Hemin, a byproduct of hemoglobin degradation, inflicts oxidative insult to cells. Following its accumulation, several proteins are recruited for heme detoxification with heme oxygenase playing the key role. Chaperones play a protective role primarily by preventing protein degradation and unfolding. They also are known to have miscellaneous secondary roles during similar situations. To discover a secondary role of chaperones during heme stress we studied the role of the chaperone HSPA8 in the detoxification of hemin. In-silico studies indicated that HSPA8 has a well-defined biophoric environment to bind hemin. Through optical difference spectroscopy, we found that HSPA8 binds hemin through its N-terminal domain with a Kd value of 5.9 ± 0.04 µM and transforms into a hemoprotein. The hemoprotein was tested for exhibiting peroxidase activity using guaiacol as substrate. The complex formed reacts with H2O2 and exhibits classical peroxidase activity with an ability to oxidize aromatic and halide substrates. HSPA8 is dose-dependently catalyzing heme polymerization through its N-terminal domain. The IR results reveal that the polymer formed exhibits structural similarities to β-hematin suggesting its covalent nature. The polymerization mechanism was tested through optical spectroscopy, spin-trap, and activity inhibition experiments. The results suggest that the polymerization occurs through a peroxidase-H2O2 system involving a one-electron transfer mechanism, and the formation of free radical and radical-radical interaction. It highlights a possible role of the HSPA8-hemin complex in exhibiting cytoprotective function during pathological conditions like malaria, sickle cell disease, etc.

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来源期刊
The Protein Journal
The Protein Journal 生物-生化与分子生物学
CiteScore
5.20
自引率
0.00%
发文量
57
审稿时长
12 months
期刊介绍: The Protein Journal (formerly the Journal of Protein Chemistry) publishes original research work on all aspects of proteins and peptides. These include studies concerned with covalent or three-dimensional structure determination (X-ray, NMR, cryoEM, EPR/ESR, optical methods, etc.), computational aspects of protein structure and function, protein folding and misfolding, assembly, genetics, evolution, proteomics, molecular biology, protein engineering, protein nanotechnology, protein purification and analysis and peptide synthesis, as well as the elucidation and interpretation of the molecular bases of biological activities of proteins and peptides. We accept original research papers, reviews, mini-reviews, hypotheses, opinion papers, and letters to the editor.
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