Department of Chemistry, Wayne State University, 5101, Cass Avenue, Detroit, Michigan 48202, USA.
Bioconjug Chem. 2012 Mar 21;23(3):650-5. doi: 10.1021/bc200694t. Epub 2012 Mar 8.
A new method was developed for site-specific modifications of liposomes by proteins via sortase A (SrtA)-mediated transpeptidation reactions. In this regard, the enhanced green fluorescent protein (eGFP) was biologically engineered to carry at its polypeptide C-terminus the LPATG motif recognized by SrtA and used as the protein donor for linking to liposomes that were decorated with phospholipids carrying a diglycine motif as the other SrtA substrate and the eGFP acceptor. Under the influence of SrtA, eGFP was efficiently attached to liposomes, as proved by analyzing the enzymatic reaction products and the resultant fluorescent liposomes. It was observed that increasing the concentration and the distance of the diglycine motif on and from the liposome surface could significantly improve the efficiency of liposome modification by proteins. It is anticipated that this strategy can be widely useful for the modification of liposomes by other proteins.
一种新方法通过 sortase A（SrtA）介导的转肽反应，实现了针对脂质体的位点特异性修饰。在这方面，对增强型绿色荧光蛋白（eGFP）进行了生物工程改造，使其在多肽 C 末端携带 SrtA 识别的 LPATG 基序，并用作与脂质体连接的蛋白质供体，脂质体上修饰有携带二甘氨酸基序的磷脂，作为另一个 SrtA 底物和 eGFP 受体。在 SrtA 的影响下，如通过分析酶反应产物和所得荧光脂质体所证明的，eGFP 被有效地连接到脂质体上。观察到增加二甘氨酸基序在脂质体表面上和从脂质体表面的距离和浓度可以显著提高蛋白质对脂质体修饰的效率。预计该策略可广泛用于其他蛋白质对脂质体的修饰。
