Sauerzapfe Birgit, Krenek Karel, Schmiedel Judith, Wakarchuk Warren W, Pelantová Helena, Kren Vladimir, Elling Lothar
Institute of Biotechnology and Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University, Germany.
Glycoconj J. 2009 Feb;26(2):141-59. doi: 10.1007/s10719-008-9172-2. Epub 2008 Aug 29.
Poly-N-acetyllactosamine (poly-LacNAc) structures have been identified as important ligands for galectin-mediated cell adhesion to extra-cellular matrix (ECM) proteins. We here present the biofunctionalization of surfaces with poly-LacNAc structures and subsequent binding of ECM glycoproteins. First, we synthesized beta-GlcNAc glycosides carrying a linker for controlled coupling onto chemically functionalized surfaces. Then we produced poly-LacNAc structures with defined lengths using human beta1,4-galactosyltransferase-1 and beta1,3-N-acetylglucosaminyltransferase from Helicobacter pylori. These compounds were also used for kinetic characterization of glycosyltransferases and lectin binding assays. A mixture of poly-LacNAc-structures covalently coupled to functionalized microtiter plates were identified for best binding to our model galectin His(6)CGL2. We further demonstrate for the first time that these poly-LacNAc surfaces are suitable for further galectin-mediated binding of the ECM glycoproteins laminin and fibronectin. This new technology should facilitate cell adhesion to biofunctionalized surfaces by imitating the natural ECM microenvironment.
多聚N-乙酰乳糖胺（poly-LacNAc）结构已被确定为半乳糖凝集素介导细胞与细胞外基质（ECM）蛋白黏附的重要配体。我们在此展示了用poly-LacNAc结构对表面进行生物功能化以及随后ECM糖蛋白的结合。首先，我们合成了带有连接子的β-葡萄糖胺糖苷，用于可控地偶联到化学功能化表面。然后，我们使用人β1,4-半乳糖基转移酶-1和幽门螺杆菌的β1,3-N-乙酰葡糖胺基转移酶制备了具有确定长度的poly-LacNAc结构。这些化合物还用于糖基转移酶的动力学表征和凝集素结合测定。已确定共价偶联到功能化微孔板上的poly-LacNAc结构混合物与我们的模型半乳糖凝集素His(6)CGL2的结合效果最佳。我们首次进一步证明，这些poly-LacNAc表面适用于半乳糖凝集素介导的ECM糖蛋白层粘连蛋白和纤连蛋白的进一步结合。这项新技术应通过模拟天然ECM微环境促进细胞黏附到生物功能化表面。
