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首页> 外文期刊>ACS nano >Interactions of amino acids and polypeptides with metal oxide nanoparticles probed by fluorescent indicator adsorption and displacement
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Interactions of amino acids and polypeptides with metal oxide nanoparticles probed by fluorescent indicator adsorption and displacement

机译:氨基酸和多肽与金属氧化物纳米粒子的相互作用(通过荧光指示剂吸附和置换)

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The adsorption of polypeptides containing an N-terminal tryptophan (Trp) residue attached to a hexa-backbone of alanine, serine, lysine, histidine, and aspartate was investigated by monitoring the fluorescence response of the Trp chromophore upon titration with metal oxide nanoparticles (MOx-NPs: CuO, Co _3O _4, TiO _2, MgO, and CeO _2). After correction for light-scattering effects, a strong static fluorescence quenching was observed upon addition of CuO and Co _3O _4 to the peptides. The interaction of MOx-NPs with the peptides was assigned to an adsorption of the peptide backbone on the nanoparticle surface. The method was refined using a derivatized amino acid, 5-fluoro-Trp (5F-Trp), which resulted in a stronger fluorescence response. The use of the fluorescent amino acid labels allowed the direct assessment of the adsorption propensities of Trp-containing peptides in dependence on the backbone, which was verified by zeta-potential measurements. Moreover, upon addition of different analytes to nanoparticles with preadsorbed Trp-containing polypeptides, adsorption propensities of the analytes were assessed by an indicator displacement strategy; that is, addition of increasing amounts of analyte resulted in a continuous fluorescence enhancement/recovery. This method afforded adsorption propensities for several analytes. The relative binding constants for the MOx-NPs, obtained from the competitive titrations, varied by more than 6 orders of magnitude for CuO (5F-TrpHis _6-NH _2 > TrpAsp _6-NH _2, TrpSer _6-NH _2 > TrpLys _6-NH _2, Trp, 5F-Trp > TrpAla _6-NH _2) but only _4 for Co _3O _4 (TrpHis _6-NH _2, TrpAsp _6-NH _2 ? TrpLys _6-NH _2, TrpAla _6-NH _2, TrpSer _6-NH _2, Trp, 5F-Trp). The study reveals that MOx-NPs adsorb biomolecular analytes with high selectivity, which has immediate implications for their applications in protein purification, drug delivery, and, potentially, for the assessment of their toxicology.
机译:通过监测金属氧化物纳米粒子(MOx)滴定后Trp发色团的荧光响应,研究了含有N末端色氨酸(Trp)残基的多肽吸附到丙氨酸,丝氨酸,赖氨酸,组氨酸和天冬氨酸的六主链上的吸附情况。 -NPs:CuO,Co _3O _4,TiO _2,MgO和CeO _2)。校正光散射效应后,在向肽中添加CuO和Co _3O _4时,观察到了强烈的静态荧光猝灭。 MOx-NP与肽的相互作用被分配为肽主链在纳米颗粒表面上的吸附。该方法使用衍生化的氨基酸5-fluoro-Trp(5F-Trp)进行了改进,从而产生了更强的荧光响应。荧光氨基酸标记的使用允许直接评估含Trp的肽根据主链的吸附倾向,这已通过Zeta电位测量得到证实。此外,在将不同的分析物添加到具有预先吸附的含Trp多肽的纳米颗粒后,通过指示剂置换策略评估了分析物的吸附倾向。也就是说,添加增加量的分析物导致连续的荧光增强/回收。这种方法提供了几种分析物的吸附倾向。从竞争性滴定获得的MOx-NP的相对结合常数对于CuO的变化超过6个数量级(5F-TrpHis _6-NH _2> TrpAsp _6-NH _2,TrpSer _6-NH _2> TrpLys _6- NH _2,Trp,5F-Trp> TrpAla _6-NH _2),但对于Co _3O _4(TrpHis _6-NH _2,TrpAsp _6-NH _2?TrpLys _6-NH _2,TrpAla _6-NH _2,TrpSer _6-只有_4 NH _2,色氨酸,5F-色氨酸)。这项研究表明,MOx-NPs可以高选择性地吸附生物分子分析物,这对其在蛋白质纯化,药物输送以及潜在地对其毒理学评估中的应用有着直接的影响。

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