Institute of Chemistry, Saint Petersburg State University, 198504 St. Petersburg, Russia.
Department of Chemical Engineering, National Taiwan University of Science and Technology, 43, Keelung Road, Section 4, Taipei 106, Taiwan.
Int J Mol Sci. 2022 Oct 16;23(20):12377. doi: 10.3390/ijms232012377.
The interactions of DNA with lysozyme in the surface layer were studied by performing infrared reflection-absorption spectroscopy (IRRAS), ellipsometry, surface tensiometry, surface dilational rheology, and atomic force microscopy (AFM). A concentrated DNA solution was injected into an aqueous subphase underneath a spread lysozyme layer. While the optical properties of the surface layer changed fast after DNA injection, the dynamic dilational surface elasticity almost did not change, thereby indicating no continuous network formation of DNA/lysozyme complexes, unlike the case of DNA interactions with a monolayer of a cationic synthetic polyelectrolyte. A relatively fast increase in optical signals after a DNA injection under a lysozyme layer indicates that DNA penetration is controlled by diffusion. At low surface pressures, the AFM images show the formation of long strands in the surface layer. Increased surface compression does not lead to the formation of a network of DNA/lysozyme aggregates as in the case of a mixed layer of DNA and synthetic polyelectrolytes, but to the appearance of some folds and ridges in the layer. The formation of more disordered aggregates is presumably a consequence of weaker interactions of lysozyme with duplex DNA and the stabilization, at the same time, of loops of unpaired nucleotides at high local lysozyme concentrations in the surface layer.
采用红外反射吸收光谱(IRRAS)、椭圆偏振法、表面张力法、表面膨胀流变学和原子力显微镜(AFM)研究了 DNA 与表面层溶菌酶的相互作用。将浓缩的 DNA 溶液注入铺展溶菌酶层下方的水性亚相中。DNA 注入后,表面层的光学性质迅速变化,而动态膨胀表面弹性几乎没有变化,这表明 DNA/溶菌酶复合物没有形成连续的网络,与 DNA 与阳离子合成聚电解质单层的相互作用不同。在溶菌酶层下注入 DNA 后,光学信号的相对快速增加表明 DNA 的穿透受扩散控制。在低表面压力下,AFM 图像显示在表面层中形成了长链。增加表面压缩不会导致 DNA/溶菌酶聚集体网络的形成,如 DNA 和合成聚电解质的混合层那样,而是导致层中出现一些褶皱和脊。更多无序聚集的形成可能是由于溶菌酶与双链 DNA 的相互作用较弱,同时在表面层中局部溶菌酶浓度较高时稳定了未配对核苷酸的环。