Pavlova Anna S, Ilyushchenko Valeriya V, Kupryushkin Maxim S, Zharkov Timofey D, Dyudeeva Evgeniya S, Bauer Irina A, Chubarov Alexey S, Pyshnyi Dmitrii V, Pyshnaya Inna A
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090, Russia.
Biochemistry (Mosc). 2023 Aug;88(8):1165-1180. doi: 10.1134/S0006297923080102.
Serum albumin is currently in the focus of biomedical research as a promising platform for the creation of multicomponent self-assembling systems due to the presence of several sites with high binding affinity of various compounds in its molecule, including lipophilic oligonucleotide conjugates. In this work, we investigated the stoichiometry of the dodecyl-containing oligonucleotides binding to bovine and human serum albumins using an electrophoretic mobility shift assay. The results indicate the formation of the albumin-oligonucleotide complexes with a stoichiometry of about 1 : (1.25 ± 0.25) under physiological-like conditions. Using atomic force microscopy, it was found that the interaction of human serum albumin with the duplex of complementary dodecyl-containing oligonucleotides resulted in the formation of circular associates with a diameter of 165.5 ± 94.3 nm and 28.9 ± 16.9 nm in height, and interaction with polydeoxyadenylic acid and dodecyl-containing oligothymidylate resulted in formation of supramolecular associates with the size of about 315.4 ± 70.9 and 188.3 ± 43.7 nm, respectively. The obtained data allow considering the dodecyl-containing oligonucleotides and albumin as potential components of the designed self-assembling systems for solving problems of molecular biology, biomedicine, and development of unique theranostics with targeted action.
血清白蛋白目前是生物医学研究的焦点，由于其分子中存在多个对各种化合物具有高结合亲和力的位点，包括亲脂性寡核苷酸缀合物，它是创建多组分自组装系统的一个有前景的平台。在这项工作中，我们使用电泳迁移率变动分析研究了含十二烷基的寡核苷酸与牛血清白蛋白和人血清白蛋白结合的化学计量。结果表明，在类似生理条件下形成了化学计量约为1:(1.25±0.25)的白蛋白-寡核苷酸复合物。使用原子力显微镜发现，人血清白蛋白与含十二烷基的互补寡核苷酸双链体的相互作用导致形成直径为165.5±94.3nm、高度为28.9±16.9nm的圆形聚集体，与聚脱氧腺苷酸和含十二烷基的寡聚胸苷酸的相互作用分别导致形成尺寸约为315.4±70.9nm和188.3±43.7nm的超分子聚集体。所获得的数据使得可以将含十二烷基的寡核苷酸和白蛋白视为用于解决分子生物学、生物医学问题以及开发具有靶向作用的独特治疗诊断学的设计自组装系统的潜在组分。
