Zheng Haifu, Du Xuezhong
Key Laboratory of Mesoscopic Chemistry (Ministry of Education), State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, People's Republic of China.
Biochim Biophys Acta. 2013 Feb;1828(2):792-800. doi: 10.1016/j.bbamem.2012.11.006. Epub 2012 Nov 14.
Imprinted monolayers provide several advantages over bulk imprinting methods. This is especially important for large templates such as proteins. Concanavalin A (Con A)-imprinted binary monolayers consisting of glycolipids with oligo(ethylene glycol) (OEG) spacers and zwitterionic phospholipids (DPPC) were constructed and investigated. The shorter phosphorylcholine (PC) headgroups with an almost flat-on orientation in the binary monolayers gave rise to reduced steric hindrance favorable to the accommodation of Con A with greater ease and facilitated the access of the OEG-linked mannose moieties for enhanced protein binding. Further enhanced binding resulted from optimized spatial rearrangement of the glycolipids at the air-water interface directed by Con A in the subphase to create bivalent binding sites and to minimize steric crowding of neighboring mannose ligands. The combination of the exposed carbohydrate ligands from biologically inert surfaces and the optimized ligand arrangement is the most reasonable solution to enhancement of protein affinity. The bivalent carbohydrate binding sites protruding from the imprinted monolayers were created to be complementary to the Con A binding pockets. This strategy generates tailor-made surfaces with enhanced protein binding and opens the possibility of controlled assembly of intellectual biomaterials and preparation of biosensors.
与本体印迹方法相比，印迹单分子层具有若干优势。这对于诸如蛋白质等大型模板尤为重要。构建并研究了由带有寡聚（乙二醇）（OEG）间隔基的糖脂和两性离子磷脂（DPPC）组成的伴刀豆球蛋白A（Con A）印迹二元单分子层。二元单分子层中具有几乎平躺取向的较短磷酰胆碱（PC）头基导致空间位阻减小，有利于更轻松地容纳Con A，并促进了与OEG连接的甘露糖部分的接触，从而增强了蛋白质结合。通过亚相中Con A引导糖脂在气-水界面进行优化的空间重排，形成二价结合位点并使相邻甘露糖配体的空间拥挤最小化，从而进一步增强了结合。来自生物惰性表面的暴露碳水化合物配体与优化的配体排列相结合，是增强蛋白质亲和力的最合理解决方案。从印迹单分子层突出的二价碳水化合物结合位点被设计成与Con A结合口袋互补。这种策略产生了具有增强蛋白质结合能力的定制表面，并为智能生物材料的可控组装和生物传感器的制备开辟了可能性。
