Belej Dominik, Jurasekova Zuzana, Nemergut Michal, Wagnieres Georges, Jancura Daniel, Huntosova Veronika
Department of Biophysics, Faculty of Science, P.J. Safarik University, Jesenna 5, 041 54 Kosice, Slovakia.
Department of Biophysics, Faculty of Science, P.J. Safarik University, Jesenna 5, 041 54 Kosice, Slovakia; Center for Interdisciplinary Biosciences, Faculty of Science, P.J. Safarik University, Jesenna 5, 041 54 Kosice, Slovakia.
J Inorg Biochem. 2017 Sep;174:37-44. doi: 10.1016/j.jinorgbio.2017.05.016. Epub 2017 May 31.
The interaction between a ruthenium - based water soluble oxygen probe ([Ru(Phen)], phen - phenanthroline) and human serum albumin (HSA) was investigated with the aim of describing the influence of HSA on the [Ru(Phen)] luminescence properties. Nowadays, several oxygen sensitive luminescent probes are used to determine the oxygen level in different compartments of living organisms. However, they can interact, depending on their hydrophilic/hydrophobic characters, with various serum proteins, and/or lipids, during their utilization for invivo oxygen measurement. Since HSA is the most abundant serum protein in most biological organisms, its presence may affect the spectral properties of the employed probes and, consequently, the determination of the oxygen concentration. Having this in mind, we have applied several spectroscopic and calorimetric techniques to study [Ru(Phen)] - HSA mixtures. Only a negligible effect of HSA on the absorption and luminescence spectra of [Ru(Phen)] was observed. In addition, differential scanning calorimetric studies showed that [Ru(Phen)] does not significantly influence HSA thermal stability. Importantly, [Ru(Phen)] retained a reliable luminescence lifetime sensitivity to the oxygen concentration in solutions supplemented with HSA and in U87 MG cancer cells. Finally, the biodistribution of [Ru(Phen)] in the presence of serum proteins in the blood stream of chick embryo's chorioallantoic membrane (CAM) was investigated. Fast [Ru(Phen)] and similar extravasations were observed in the presence or absence of CAM-serum. We can conclude that HSA-[Ru(Phen)] complex interaction does not significantly influence the potential of [Ru(Phen)] to be a suitable candidate for a reliable oxygen probe in living organisms.
研究了一种基于钌的水溶性氧探针（[Ru（Phen）]，Phen为邻菲罗啉）与人血清白蛋白（HSA）之间的相互作用，目的是描述HSA对[Ru（Phen）]发光特性的影响。如今，几种氧敏感发光探针被用于测定生物体不同部位的氧水平。然而，在用于体内氧测量的过程中，它们会根据其亲水/疏水特性与各种血清蛋白和/或脂质相互作用。由于HSA是大多数生物体内最丰富的血清蛋白，其存在可能会影响所使用探针的光谱特性，进而影响氧浓度的测定。考虑到这一点，我们应用了几种光谱和量热技术来研究[Ru（Phen）]-HSA混合物。仅观察到HSA对[Ru（Phen）]的吸收光谱和发光光谱有可忽略不计的影响。此外，差示扫描量热研究表明，[Ru（Phen）]对HSA的热稳定性没有显著影响。重要的是，[Ru（Phen）]在添加了HSA的溶液和U87 MG癌细胞中对氧浓度保持可靠的发光寿命敏感性。最后，研究了[Ru（Phen）]在鸡胚绒毛尿囊膜（CAM）血流中存在血清蛋白时的生物分布。在有或没有CAM血清的情况下，均观察到[Ru（Phen）]快速且相似的外渗现象。我们可以得出结论，HSA-[Ru（Phen）]复合物的相互作用不会显著影响[Ru（Phen）]成为生物体内可靠氧探针的潜力。
