Morales-García Ana L, Walton Rachel, Blakeman Jamie T, Banwart Steven A, Harding John H, Geoghegan Mark, Freeman Colin L, Rolfe Stephen A
Department of Physics and Astronomy, The University of Sheffield, Hounsfield Road, Sheffield S3 7RH, U.K.
Department of Animal and Plant Sciences, The University of Sheffield, Western Bank, Sheffield S10 2TN, U.K.
Langmuir. 2021 Aug 4;37(32):9838-50. doi: 10.1021/acs.langmuir.1c01410.
Attachment assays of a isolate to fused silica slides showed that treatment with DNaseI significantly inhibited cellular adsorption, which was restored upon DNA treatment. These assays confirmed the important role of extracellular DNA (eDNA) adsorption to a surface. To investigate the eDNA adsorption mechanism, single-molecule force spectroscopy (SMFS) was used to measure the adsorption of eDNA to silicon surfaces in the presence of different concentrations of sodium and calcium ions. SMFS reveals that the work of adhesion required to remove calcium-bound eDNA from the silicon oxide surface is substantially greater than that for sodium. Molecular dynamics simulations were also performed, and here, it was shown that the energy gain in eDNA adsorption to a silicon oxide surface in the presence of calcium ions is small and much less than that in the presence of sodium. The simulations show that the length scales involved in eDNA adsorption are less in the presence of sodium ions than those in the presence of calcium. In the presence of calcium, eDNA is pushed above the surface cations, whereas in the presence of sodium ions, short-range interactions with the surface dominate. Moreover, SMFS data show that increasing [Ca] from 1 to 10 mM increases the adsorption of the cations to the silicon oxide surface and consequently enhances the Stern layer, which in turn increases the length scale associated with eDNA adsorption.
一株分离株与熔融石英载玻片的附着试验表明,用DNA酶I处理可显著抑制细胞吸附,而DNA处理后吸附得以恢复。这些试验证实了细胞外DNA(eDNA)吸附到表面的重要作用。为了研究eDNA的吸附机制,采用单分子力谱(SMFS)在不同浓度的钠离子和钙离子存在下测量eDNA对硅表面的吸附。SMFS显示,从氧化硅表面去除钙结合的eDNA所需的粘附功远大于去除钠结合的eDNA所需的粘附功。还进行了分子动力学模拟,结果表明,在钙离子存在下eDNA吸附到氧化硅表面的能量增益很小,远小于在钠离子存在下的能量增益。模拟结果表明,钠离子存在下eDNA吸附涉及的长度尺度小于钙离子存在下的长度尺度。在钙离子存在下,eDNA被推到表面阳离子上方,而在钠离子存在下,与表面的短程相互作用占主导。此外,SMFS数据表明,将[Ca]从1 mM增加到10 mM会增加阳离子对氧化硅表面的吸附,从而增强斯特恩层,进而增加与eDNA吸附相关的长度尺度。