Worcester Polytechnic Institute, 100 Institute Road, Worcester, Massachusetts 01609, United States.
Langmuir. 2010 Dec 21;26(24):18884-92. doi: 10.1021/la103333c. Epub 2010 Nov 18.
Human red blood cell acetylcholinesterase was incorporated into planar lipid membranes deposited on alkanethiol self-assembled monolayers (SAMs) on gold substrates. Activity of the protein in the membrane was detected with a standard photometric assay and was determined to be similar to the protein in detergent solution or incorporated in lipid vesicles. Monolayer and bilayer lipid membranes were generated by fusing liposomes to hydrophobic and hydrophilic SAMs, respectively. Liposomes were formed by the injection method using the lipid dimyristoylphosphatidylcholine (DMPC). The formation of alkanethiol SAMs and lipid monolayers on SAMs was confirmed by sessile drop goniometry, ellipsometry, and electrochemical impedance spectroscopy. In this work, we report acetylcholinesterase immobilization in lipid membranes deposited on SAMs formed on the gold surface and compare its activity to enzyme in solution.
人红细胞乙酰胆碱酯酶被整合到在金基底上的烷硫醇自组装单层(SAM)上沉积的平面脂质膜中。用标准分光光度法检测蛋白质在膜中的活性,发现其与去污剂溶液中的蛋白质或脂质囊泡中的蛋白质相似。通过将脂质体分别融合到疏水性和亲水性 SAM 上来生成单层和双层脂质膜。脂质体通过使用脂质二肉豆蔻酰磷脂酰胆碱(DMPC)的注射方法形成。通过 sessile drop 测角法、椭圆偏振法和电化学阻抗谱法证实了烷硫醇 SAM 和 SAM 上的脂质单层的形成。在这项工作中,我们报告了在金表面上形成的 SAM 上沉积的脂质膜中乙酰胆碱酯酶的固定化,并将其活性与溶液中的酶进行了比较。