Mohd Shahnawaz Khan, Md Tabish Rehman, Gouse M Shaik, Abdulaziz Mohammed Alamri, Mohamed F AlAjmi, Mohammed Arshad, Majed S Alokail
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The loss of tryptophan fluorescence in HSA suggested significant structural alterations, particularly around aromatic residues. Far UV-CD analysis demonstrated disruptions in HSA's secondary structure, with a notable reduction in α-helical structures at pH 7.4. At pH 3.5, Azorubine induced even more extensive perturbations, resulting in a random coil conformation at higher azorubine concentrations. The study also investigated aggregation phenomena through turbidity measurements, RLS analysis, and TEM imaging. At pH 3.5, larger insoluble aggregates formed, while at pH 7.4, only conformational changes occurred without aggregate formation. Cytotoxicity assessments on neuroblastoma (SH-SY5Y) cells highlighted the concentration-dependent toxicity of albumin aggregates. Molecular dynamics simulations reaffirmed the stable interaction between azorubine and HSA. This research provides valuable insights into the mechanisms by which azorubine influences protein conformations. 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引用次数: 0
摘要
蛋白质和肽的错误折叠是与阿尔茨海默病和帕金森病等疾病相关的病理性聚集体和原纤维形成的核心因素。因此,有必要了解食品添加剂,特别是偶氮吡啶,如何影响蛋白质结构及其诱导聚集的能力。本研究以人血清白蛋白(HSA)为模型蛋白,研究了偶氮吡啶(一种常见的食品和饮料着色剂)对其结合和构象的影响。研究发现,在生理(pH 7.4)和酸性(pH 3.5)条件下,偶氮吡啶都能破坏HSA的构象。HSA中色氨酸荧光的缺失表明了显著的结构改变,特别是在芳香残基周围。远紫外- cd分析表明,在pH值为7.4时,HSA的二级结构被破坏,α-螺旋结构明显减少。在pH为3.5时,偶氮吡啶引起更广泛的扰动,在较高的偶氮吡啶浓度下导致随机线圈构象。该研究还通过浊度测量、RLS分析和TEM成像研究了聚集现象。pH为3.5时,形成较大的不溶性聚集体,而pH为7.4时,仅发生构象变化,未形成聚集体。神经母细胞瘤(SH-SY5Y)细胞的细胞毒性评估强调了白蛋白聚集体的浓度依赖性毒性。分子动力学模拟证实了偶氮吡啶与HSA之间稳定的相互作用。这项研究为偶氮嘌呤影响蛋白质构象的机制提供了有价值的见解。为了进一步加深我们对这一领域的理解,未来的几个方向可以考虑,如探索其他蛋白质,研究剂量-反应关系,获得机制见解,生物学相关性,毒性评估,确定替代食用色素,以及缓解策略,以防止偶氮吡啶对血清蛋白的不良影响。由Ramaswamy H. Sarma传达。
Aggregation and cytotoxicity of food additive dye (Azorubine)-albumin adducts: a multi-spectroscopic, microscopic and computational analysis.
Protein and peptide misfolding is a central factor in the formation of pathological aggregates and fibrils linked to disorders like Alzheimer's and Parkinson's diseases. Therefore, it's essential to understand how food additives, particularly Azorubine, affect protein structures and their ability to induce aggregation. In this study, human serum albumin (HSA) was used as a model protein to investigate the binding and conformational changes caused by azorubine, a common food and drink colorant. The research revealed that azorubine destabilized the conformation of HSA at both physiological (pH 7.4) and acidic (pH 3.5) conditions. The loss of tryptophan fluorescence in HSA suggested significant structural alterations, particularly around aromatic residues. Far UV-CD analysis demonstrated disruptions in HSA's secondary structure, with a notable reduction in α-helical structures at pH 7.4. At pH 3.5, Azorubine induced even more extensive perturbations, resulting in a random coil conformation at higher azorubine concentrations. The study also investigated aggregation phenomena through turbidity measurements, RLS analysis, and TEM imaging. At pH 3.5, larger insoluble aggregates formed, while at pH 7.4, only conformational changes occurred without aggregate formation. Cytotoxicity assessments on neuroblastoma (SH-SY5Y) cells highlighted the concentration-dependent toxicity of albumin aggregates. Molecular dynamics simulations reaffirmed the stable interaction between azorubine and HSA. This research provides valuable insights into the mechanisms by which azorubine influences protein conformations. To further advance our understanding and contribute to the broader knowledge in this area, several future directions can be considered such as exploring other proteins, studying dose-response relationship, gaining mechanistic insights, biological relevance, toxicity assessment, identifying alternative food colorants, and mitigation strategies to prevent adverse effects of azorubine on serum proteins.Communicated by Ramaswamy H. Sarma.
期刊介绍:
The Journal of Biomolecular Structure and Dynamics welcomes manuscripts on biological structure, dynamics, interactions and expression. The Journal is one of the leading publications in high end computational science, atomic structural biology, bioinformatics, virtual drug design, genomics and biological networks.