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Modular Genetic Code Expansion Platform and PISA Yield Well-Defined Protein-Polymer Assemblies
Biomacromolecules ( IF 5.5 ) Pub Date : 2020-11-10 , DOI: 10.1021/acs.biomac.0c01225
Dominic J. Rucco 1 , Brooke E. Barnes 1 , John B. Garrison 1 , Brent S. Sumerlin 1 , Daniel A. Savin 1
Affiliation  

We present a modular platform from which biohybrid protein-polymer nanostructures can be generated in a straightforward and facile manner. Specifically, an aqueous polymerization-induced self-assembly (PISA) AB block copolymerization system was derived from a mutant superfolder green fluorescent protein (sfGFP) as the solvophilic, stabilizing A block. By genetically encoding sfGFP with an isobutyryl bromide functionality, we grafted a quintessential atom-transfer radical polymerization initiation site with hydroxypropyl methacrylate (HPMA) to form the solvophobic B block. Monitoring nanostructure formation using dynamic light scattering, gel permeation chromatography, and transmission electron microscopy revealed uniform micellar morphologies. The radii of the micelles increased with increasing HPMA block length, resulting in nanoparticle sizes ranging from 15 to 48 nm. Solvophilic stabilization afforded by the encoded sfGFP makes this an ideal PISA initiator, and we posit this platform has potential for generating complex biohybrid nanostructures for other protein–polymer systems.

中文翻译:

模块化遗传密码扩展平台和PISA可产生定义良好的蛋白质-聚合物组件

我们提出了一个模块化的平台,可以从该平台以简单易行的方式生成生物杂交蛋白-聚合物纳米结构。具体而言,水性聚合诱导的自组装(PISA)AB嵌段共聚体系衍生自突变超文件夹绿色荧光蛋白(sfGFP),为亲溶剂性,稳定化的A嵌段。通过遗传编码具有异丁酰基溴官能团的sfGFP,我们将典型的原子转移自由基聚合起始位点与甲基丙烯酸羟丙酯(HPMA)接枝形成疏溶剂性B嵌段。使用动态光散射,凝胶渗透色谱和透射电子显微镜监测纳米结构的形成显示出均匀的胶束形态。胶束半径随HPMA嵌段长度的增加而增加,产生的纳米粒子尺寸范围为15至48 nm。编码的sfGFP提供的亲溶剂稳定作用使其成为理想的PISA引发剂,并且我们认为该平台具有为其他蛋白质-聚合物系统产生复杂的生物杂交纳米结构的潜力。
更新日期:2020-12-14
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