Key Laboratory of Functional Polymer Materials, Ministry of Education, College of Chemistry, Collaborative Innovation Center of Chemical, Science and Engineering (Tianjin), Nankai University, P.R. China.
Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Sciences, Nankai University, P.R. China.
Chemistry. 2017 Mar 8;23(14):3366-3374. doi: 10.1002/chem.201604843. Epub 2017 Feb 7.
The synthesis of polymer-protein nanostructures opens up a new avenue for the development of new biomaterials. In this research, covalently connected polymer-protein nanostructures were fabricated through a reactive self-assembly approach. Poly(tert-butyl methacrylate-co-pyridyl disulfide methacrylamide) (PtBMA-co-PPDSMA) was synthesized by reversible addition fragmentation chain transfer (RAFT) polymerization. Covalently connected nanostructures (CCNs) with hydrophobic polymer cores and hydrophilic protein coronae were prepared by adding solutions of PtBMA-co-PPDSMA/DMF to aqueous solutions of bovine serum albumin (BSA). The thiol-disulfide exchange reaction between pyridyl disulfide groups on the polymer chains and thiol groups on the protein molecules plays a key role in the fabrication of CCNs. The self-assembly process was investigated by dynamic light scattering (DLS) and stopped-flow techniques. DLS results indicated that the sizes of the CCNs were determined by the initial polymer concentration, the BSA concentration, and the average number of thiol groups on BSA molecules. TEM and sodium dodecyl sulfate polyacrylamide gel electrophoresis were used to analyze the nanostructures. Far-UV circular dichroism results demonstrated that the original folded conformations of BSA molecules were basically maintained in the reactive self-assembly process. Compared with native BSA, the secondary structure and conformation change of coronal BSA induced by urea or thermal treatment were remarkably suppressed. The cytotoxicity assays demonstrated that the CCNs were essentially nontoxic to Hela and COS-7 cells.
聚合物-蛋白质纳米结构的合成为新型生物材料的发展开辟了新途径。在这项研究中，通过反应性自组装方法制备了共价连接的聚合物-蛋白质纳米结构。通过可逆加成-断裂链转移（RAFT）聚合合成了聚（叔丁基甲基丙烯酸酯-co-吡啶二硫代甲酰胺甲基丙烯酰胺）（PtBMA-co-PPDSMA）。通过将 PtBMA-co-PPDSMA/DMF 溶液添加到牛血清白蛋白（BSA）的水溶液中，制备了具有疏水性聚合物核和亲水性蛋白质冠状物的共价连接纳米结构（CCN）。聚合物链上的吡啶二硫代基团和蛋白质分子上的巯基之间的硫代-二硫交换反应在 CCN 的制备中起着关键作用。通过动态光散射（DLS）和停流技术研究了自组装过程。DLS 结果表明，CCN 的尺寸取决于初始聚合物浓度、BSA 浓度和 BSA 分子上的巯基数。TEM 和十二烷基硫酸钠聚丙烯酰胺凝胶电泳用于分析纳米结构。远紫外圆二色性结果表明，BSA 分子的原始折叠构象在反应性自组装过程中基本保持不变。与天然 BSA 相比，冠状 BSA 的二级结构和构象变化在尿素或热处理下受到显著抑制。细胞毒性测定表明，CCN 对 Hela 和 COS-7 细胞基本无毒。
