Institute of Physics CAS, Na Slovance 1999/2, 182 21 Prague, Czech Republic.
Department of Biochemistry, Faculty of Science, Masaryk University, Kamenice 5, 625 00 Brno, Czech Republic.
Biomolecules. 2020 Aug 5;10(8):1146. doi: 10.3390/biom10081146.
Cells are continuously sensing their microenvironment and subsequently respond to different physicochemical cues by the activation or inhibition of different signaling pathways. To study a very complex cellular response, it is necessary to diminish background environmental influences and highlight the particular event. However, surface-driven nonspecific interactions of the abundant biomolecules from the environment influence the targeted cell response significantly. Yes-associated protein (YAP) translocation may serve as a marker of human hepatocellular carcinoma (Huh7) cell responses to the extracellular matrix and surface-mediated stresses. Here, we propose a platform of tunable functionable antifouling poly(carboxybetain) (pCB)-based brushes to achieve a molecularly clean background for studying arginine, glycine, and aspartic acid (RGD)-induced YAP-connected mechanotransduction. Using two different sets of RGD-functionalized zwitterionic antifouling coatings with varying compositions of the antifouling layer, a clear correlation of YAP distribution with RGD functionalization concentrations was observed. On the other hand, commonly used surface passivation by the oligo(ethylene glycol)-based self-assembled monolayer (SAM) shows no potential to induce dependency of the YAP distribution on RGD concentrations. The results indicate that the antifouling background is a crucial component of surface-based cellular response studies, and pCB-based zwitterionic antifouling brush architectures may serve as a potential next-generation easily functionable surface platform for the monitoring and quantification of cellular processes.
细胞不断感知其微环境，并通过激活或抑制不同的信号通路来对不同的理化线索做出反应。为了研究非常复杂的细胞反应，有必要减少背景环境的影响，突出特定的事件。然而，来自环境的丰富生物分子的表面驱动的非特异性相互作用会显著影响靶细胞的反应。Yes 相关蛋白（YAP）易位可以作为人肝癌细胞（Huh7）对细胞外基质和表面介导的应激反应的标志物。在这里，我们提出了一种可调功能的抗污聚（羧基甜菜碱）（pCB）基刷的平台，以实现分子清洁的背景，用于研究精氨酸、甘氨酸和天冬氨酸（RGD）诱导的 YAP 连接的机械转导。使用两组具有不同抗污层组成的 RGD 功能化两性离子抗污涂层，观察到 YAP 分布与 RGD 功能化浓度之间存在明显的相关性。另一方面，常用的基于寡聚乙二醇的自组装单分子层（SAM）表面钝化没有潜力诱导 YAP 分布对 RGD 浓度的依赖性。结果表明，抗污背景是基于表面的细胞反应研究的关键组成部分，pCB 基两性离子抗污刷结构可以作为一种潜在的下一代易于功能化的表面平台，用于监测和量化细胞过程。
