Zapata Angela, Nguyen Mai-Loan, Ling Caleb, Rogers Jacqueline, Domiano Sangeetha, Hayzelden Clive, Wheeler Korin E
Department of Chemistry & Biochemistry, Santa Clara University, Santa Clara, CA 95053, USA.
Department of Biology, San Francisco State University, San Francisco, CA 94132, USA.
Nanoscale Adv. 2020 Jun 1;2(6):2429-2440. doi: 10.1039/C9NA00793H. Epub 2020 Apr 21.
In living systems, the biomolecules that coat nanoparticles (NPs) alter the NP biological identity and response. Although some biomolecules are more effective in mediating NP stability or biological fate, it is difficult to monitor an individual biomolecule within the complexity of the biota. To understand the dependence of protein-NP interactions on common variations in blood, we have evaluated binding between silica NPs and a model gamma-fibrinogen (GF) peptide. Fibrinogen is commonly identified within the protein corona fingerprint of human serum, but its abundance on the NP varies. To assess the relative importance of human serum and solution conditions, GF peptide and silica NP interactions were evaluated with and without serum across pH, NaCl concentrations, and glucose concentrations. Initial evaluation of the GF peptide and silica NP complexes using circular dichroism and dynamic light scattering show little change in the secondary structure of the peptide and no significant agglomeration of NPs, suggesting peptide-NP complexes are stable across study conditions. Fluorescence anisotropy was used to monitor GF peptide-NP binding. Both with and without serum, binding constants for the gamma-fibrinogen peptide vary significantly upon addition of diluted HS (1:500) and 29 mM sodium chloride. Yet, results indicated that gamma-fibrinogen binding interactions with silica NPs are comparatively insensitive to physiologically relevant pH changes and dramatic increases in glucose concentrations. Results highlight the importance of blood chemistries, which vary across individuals and disease states, in mediating protein corona formation.
在生命系统中,包覆纳米颗粒(NP)的生物分子会改变NP的生物学特性和反应。尽管一些生物分子在介导NP稳定性或生物学命运方面更有效,但在生物群的复杂性中监测单个生物分子却很困难。为了了解蛋白质-NP相互作用对血液中常见变化的依赖性,我们评估了二氧化硅NP与模型γ-纤维蛋白原(GF)肽之间的结合。纤维蛋白原通常在人血清的蛋白质冠指纹中被识别,但其在NP上的丰度有所不同。为了评估人血清和溶液条件的相对重要性,在有和没有血清的情况下,分别在不同pH值、NaCl浓度和葡萄糖浓度下评估了GF肽与二氧化硅NP的相互作用。使用圆二色性和动态光散射对GF肽和二氧化硅NP复合物进行的初步评估表明,肽的二级结构几乎没有变化,NP也没有明显的团聚,这表明肽-NP复合物在整个研究条件下都是稳定的。荧光各向异性用于监测GF肽与NP的结合。无论有无血清,添加稀释的人血清(1:500)和29 mM氯化钠后,γ-纤维蛋白原肽的结合常数都会发生显著变化。然而,结果表明,γ-纤维蛋白原与二氧化硅NP的结合相互作用对生理相关的pH变化和葡萄糖浓度的显著增加相对不敏感。结果突出了血液化学成分在介导蛋白质冠形成中的重要性,而血液化学成分在个体和疾病状态之间存在差异。