Miotto Giovanni, Magro Massimiliano, Terzo Milo, Zaccarin Mattia, Da Dalt Laura, Bonaiuto Emanuela, Baratella Davide, Gabai Gianfranco, Vianello Fabio
Department of Molecular Medicine, University of Padua, Italy; Proteomics Facility, Azienda Ospedaliera di Padova, University of Padua, Italy.
Department of Comparative Biomedicine and Food Science, University of Padua, Italy; Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry and Experimental Physics, Faculty of Science, Palacky University, 17. Listopadu 1192/12, 771 46 Olomouc, Czech Republic.
Colloids Surf B Biointerfaces. 2016 Apr 1;140:40-49. doi: 10.1016/j.colsurfb.2015.11.043. Epub 2015 Dec 19.
Proteome modifications in a biological fluid can potentially indicate the occurrence of pathologies, even if the identification of a proteome fingerprint correlated to a specific disease represents a very difficult task. When a nanomaterial is introduced into a biological fluid, macromolecules compete to form a protein corona on the nanoparticle surface, and depending on the specific proteome, different patterns of proteins will form the final protein corona shell depending on their affinity for the nanoparticle surface. Novel surface active maghemite nanoparticles (SAMNs) display a remarkable selectivity toward protein corona formation, and they are able to concentrate proteins and peptides presenting high affinities for their surface even if they are present in very low amounts. Thus, SAMNs may confer visibility to hidden biomarkers correlated to the occurrence of a pathology. In the present report, SAMNs were introduced into milk samples from healthy cows and from animals affected by mastitis, and the selectively bound protein corona shell was easily analyzed and quantified by gel electrophoresis and characterized by mass spectrometry. Upon incubation in mastitic milk, SAMNs were able to selectively bind αs2-casein fragments containing the FALPQYLK sequence, as part of the larger casocidin-1 peptide with strong antibacterial activity, which were not present in healthy samples. Thus, SAMNs can be used as a future candidate for the rapid diagnosis of mastitis in bovine milk. The present report proposes protein competition for SAMN protein corona formation as a means of mirroring proteome modifications. Thus, the selected protein shell on the nanoparticles results in a fingerprint of the specific pathology.
生物流体中的蛋白质组修饰可能预示着疾病的发生，即便确定与特定疾病相关的蛋白质组指纹是一项极具挑战性的任务。当一种纳米材料被引入生物流体中时，大分子会竞相在纳米颗粒表面形成蛋白质冠层，并且根据特定的蛋白质组，不同的蛋白质模式将依据它们对纳米颗粒表面的亲和力形成最终的蛋白质冠层外壳。新型表面活性磁赤铁矿纳米颗粒（SAMNs）对蛋白质冠层的形成表现出显著的选择性，并且它们能够富集那些即便含量极低但对其表面具有高亲和力的蛋白质和肽。因此，SAMNs可能会使与疾病发生相关的隐藏生物标志物显现出来。在本报告中，SAMNs被引入来自健康奶牛和患乳腺炎奶牛的牛奶样本中，通过凝胶电泳能够轻松地分析和定量选择性结合的蛋白质冠层外壳，并通过质谱对其进行表征。在乳腺炎牛奶中孵育后，SAMNs能够选择性地结合含有FALPQYLK序列的αs2-酪蛋白片段，该片段是具有强抗菌活性的较大的酪蛋白抗微生物肽-1的一部分，而在健康样本中不存在。因此，SAMNs可作为未来快速诊断牛奶中乳腺炎的候选物。本报告提出蛋白质竞争形成SAMN蛋白质冠层是反映蛋白质组修饰的一种方式。因此，纳米颗粒上选定的蛋白质外壳形成了特定疾病的指纹。
