Institute of Organic and Biomolecular Chemistry, University of Göttingen, Göttingen, Germany.
Institute of Physical Chemistry, University of Göttingen, Göttingen, Germany.
Chemistry. 2023 Jul 11;29(39):e202203904. doi: 10.1002/chem.202203904. Epub 2023 May 31.
Cell adhesion molecules are crucial for a variety of biological processes, including wound healing, barrier formation and tissue homeostasis. One of them is E-cadherin which is generally found at adherent junctions between epithelial cells. To identify this molecule on the surface of cells, E-cadherin mimetic peptides with a critical amino acid sequence of HAV (histidine-alanine-valine) were synthesized and attached to solid-supported membranes covering colloidal probes. Two different functionalization strategies were established, one based on the complexation of DOGS-NTA(Ni) with a polyhistidine-tagged HAV-peptide and the other one relying on the formation of a HAV-lipopeptide using in situ maleimide-thiol coupling. Binding studies were performed to verify the ability of the peptides to attach to the membrane surface. Compared to the non-covalent attachment via the His-tag, we achieved a higher yield by lipopeptide formation. Colloidal probes functionalized with HAV-peptides were employed to measure the presence of E-cadherins on living cells either using video particle tracking or force spectroscopy. Here, human HaCaT cells were examined confirming the specific interaction of the HAV-peptide with the E-cadherin of the cells. Statistical methods were also used to determine the number of single-bond ruptures and the force of a single bond. These findings may be essential for the development of novel biosynthetic materials given their potential to become increasingly relevant in medical applications.
细胞黏附分子对于多种生物学过程至关重要，包括伤口愈合、屏障形成和组织稳态。其中之一是 E-钙黏蛋白，它通常存在于上皮细胞之间的黏附连接中。为了在细胞表面识别这种分子，合成了具有 HAV（组氨酸-丙氨酸-缬氨酸）关键氨基酸序列的 E-钙黏蛋白模拟肽，并将其附着在覆盖胶体探针的固载膜上。建立了两种不同的功能化策略，一种基于 DOGS-NTA（Ni）与多组氨酸标记的 HAV-肽的络合，另一种依赖于使用原位马来酰亚胺-巯基偶联形成 HAV-脂肽。进行了结合研究以验证肽附着到膜表面的能力。与通过 His 标签的非共价附着相比，通过脂肽形成实现了更高的产率。使用视频粒子跟踪或力谱学，用人 HaCaT 细胞进行了研究，证实了 HAV-肽与细胞中的 E-钙黏蛋白的特异性相互作用。还使用统计方法来确定单键断裂的数量和单个键的力。鉴于它们在医学应用中可能变得越来越重要，这些发现对于新型生物合成材料的开发可能至关重要。
