Doltchinkova Virjinia, Kitanova Meglena, Nikolov Rumen, Stoyanova-Ivanova Angelina, Petkov Ognyan, Dikova Yoana, Vitkova Victoria
Department of Biophysics and Radiobiology, Faculty of Biology, Sofia University "St. Kliment Ohridski", 8 Dragan Tzankov blvd., 1164 Sofia, Bulgaria.
Department of Genetics, Faculty of Biology, Sofia University "St. Kliment Ohridski", 8 Dragan Tzankov blvd., 1164 Sofia, Bulgaria.
Membranes (Basel). 2023 Feb 27;13(3):281. doi: 10.3390/membranes13030281.
Pooled Immunoglobulin G (IgG), hematin and the membrane-disruptive amphipathic peptide melittin have received attention as powerful biomacromolecules for biomedical and pharmacology applications. Their action on surface properties, oxidation status and epifluorescence properties measured in vitro provide useful information about the functional activity of upper biomacromolecules in erythrocytes in vivo. The hemolysis of erythrocyte membranes, as well as changes in hematocrit and the morphology of erythrocytes, was investigated here via fluorescence microscopy using FITC-concanavalin A binding to cells. The effect of melittin on the membrane capacitance and resistance of model lipid bilayers was probed via electrochemical impedance spectroscopy. Lipid bilayer capacitance was higher in the presence of 0.10 g/L melittin compared to that in the control, which is likely related to bilayer thinning and alterations of the dielectric permittivity of melittin-treated membranes. The biomolecule interactions with red blood cells were probed in physiological media in which the surface of erythrocyte membranes was negatively charged. Surface parameters of erythrocytes are reported upon IgG/hematin and IgG/melittin treatment. Pooled IgG in the presence of melittin, preincubated IgG/hematin preparations promoted a significant decrease in the electrokinetic potential of erythrocytes (Rh-positive). A malondialdehyde (MDA) assay revealed a high rate of lipid peroxidation in erythrocytes treated with IgG/hematin or IgG/melittin preparations. This finding might be a result of pooled IgG interactions with the hematin molecule and the subsequent conformational changes in the protein molecule altering the electrokinetic properties of the erythrocyte membrane related to the Rh group type of erythrocytes. The pooled IgG and hematin are reported to have important consequences for the biophysical understanding of the immunopathological mechanisms of inflammatory, autoimmune and antibody-mediated pathological processes.
聚合免疫球蛋白G（IgG）、血红素和具有膜破坏作用的两亲性肽蜂毒素作为用于生物医学和药理学应用的强大生物大分子受到了关注。它们在体外对表面性质、氧化状态和落射荧光性质的作用，为体内红细胞中上层生物大分子的功能活性提供了有用信息。本文通过使用异硫氰酸荧光素标记的伴刀豆球蛋白A与细胞结合的荧光显微镜，研究了红细胞膜的溶血情况以及血细胞比容和红细胞形态的变化。通过电化学阻抗谱探究了蜂毒素对模型脂质双层膜电容和电阻的影响。与对照组相比，在存在0.10 g/L蜂毒素的情况下，脂质双层膜电容更高，这可能与双层膜变薄以及蜂毒素处理的膜的介电常数改变有关。在红细胞膜表面带负电荷的生理介质中探究了生物分子与红细胞的相互作用。报告了IgG/血红素和IgG/蜂毒素处理后红细胞的表面参数。在存在蜂毒素的情况下预孵育的IgG/血红素制剂，使聚合IgG导致红细胞（Rh阳性）的电动电位显著降低。丙二醛（MDA）测定显示，用IgG/血红素或IgG/蜂毒素制剂处理的红细胞中脂质过氧化率很高。这一发现可能是由于聚合IgG与血红素分子相互作用，以及随后蛋白质分子的构象变化改变了与红细胞Rh血型相关的红细胞膜的电动性质。据报道，聚合IgG和血红素对于从生物物理学角度理解炎症、自身免疫和抗体介导的病理过程的免疫病理机制具有重要意义。
