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Interaction of TiO2 nanoparticles with proteins from aquatic organisms: the case of gill mucus from blue mussel

机译:TiO2纳米粒子与水生生物蛋白的相互作用:蓝贻贝鳃粘液的情况

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To better understand the mechanisms of TiO2 nanoparticle (NP) uptake and toxicity in aquatic organisms, we investigated the interaction of NPs with the proteins found in gill mucus from blue mussels. Mucus is secreted by many aquatic organisms and is often their first line of defense against pathogens, xenobiotics, and other sources of environmental stress. Here, five TiO2 NPs and one SiO2 NP were incubated with gill mucus and run out on a one-dimensional polyacrylamide gel for a comparative qualitative analysis of the free proteins in the mucosal solution and the proteins bound to NPs. We then used nanoscale liquid chromatography coupled with tandem mass spectrometry to identify proteins of interest. Our data demonstrated dissimilar protein profiles between the crude mucosal solution and proteins adsorbed on NPs. In particular, extrapallial protein (EP), one of the most abundant mucus proteins, was absent from the adsorbed proteins. After thermal denaturation experiments, this absence was attributed to the EP content in aromatic amino acids that prevents protein unfolding and thus adsorption on the NP. Moreover, although the majority of the protein corona was qualitatively similar across the NPs tested here (SiO2 and TiO2), a few proteins in the corona showed a specific recruitment pattern according to the NP oxide (TiO2 vs SiO2) or crystal structure (anatase TiO2 vs rutile TiO2). Therefore, protein adsorption may vary with the type of NP.
机译:为了更好地了解水生生物中TiO2纳米粒子(NP)摄取和毒性的机制,我们研究了NP与来自蓝贻贝的鳃粘液中发现的蛋白质的相互作用。粘液被许多水生生物分泌,往往是对病原体,仇外病毒和其他环境压力来源的第一道防护。这里,将五个TiO 2 NPS和一个SiO 2 NP与鳃粘液一起温育,并在一维聚丙烯酰胺凝胶上耗尽,用于对粘膜溶液中的游离蛋白质和结合NPS结合的蛋白质的比较定性分析。然后使用纳米级液相色谱法与串联质谱耦合,以鉴定感兴趣的蛋白质。我们的数据在粗粘膜溶液和吸附在NPS上的蛋白质之间显示不同的蛋白质谱。特别是,吸附的蛋白质中不存在挤出蛋白质(EP),其中含有最丰富的粘液蛋白之一。在热变性实验后,这种不存在归因于芳族氨基酸中的EP含量,其防止蛋白质展开并因此吸附在NP上。此外,尽管在这里测试的NPS(SiO 2和TiO 2)上大多数蛋白质电晕定性相似,但电晕中的几种蛋白质显示了根据NP氧化物(TiO 2 Vs SiO 2)或晶体结构(Anatase TiO2)的特定募集图案VS金红石TiO2)。因此,蛋白质吸附可以随NP的类型而变化。

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