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Improved Ordering in Low Molecular Weight Protein–Polymer Conjugates Through Oligomerization of the Protein Block
Biomacromolecules ( IF 5.5 ) Pub Date : 2018-08-22 00:00:00 , DOI: 10.1021/acs.biomac.8b00928
Justin M. Paloni 1 , Eric A. Miller 1 , Hadley D. Sikes 1 , Bradley D. Olsen 1
Affiliation  

The self-assembly of protein–polymer conjugates incorporating oligomers of a small, engineered high-affinity binding protein, rcSso7d.SA, is studied to determine the effect of protein oligomerization on nanoscale ordering. Oligomerization enables a systematic increase in the protein molar mass without changing its overall folded structure, leading to a higher driving force for self-assembly into well-ordered structures. Though conjugates of monomeric rcSso7d.SA are found to only exist in disordered states, oligomers of this protein linked to a poly(N-isopropylacrylamide) (PNIPAM) block self-assemble into lamellar nanostructures. Conjugates of trimeric and tetrameric rcSso7d.SA are observed to produce the strongest ordering in concentrated solution, displaying birefringent lamellae at concentrations as low as 40 wt %. In highly concentrated solution, the oligomeric rcSso7d.SA-PNIPAM block copolymers exhibit ordering and domain spacing trends atypical from that of most block copolymers. Fluorescent binding assays indicate that oligomerized protein blocks retain binding functionality and exhibit limits of detection up to three times lower than that of surface-immobilized protein sensors. Therefore, oligomerization of the protein block in these block copolymers serves as an effective method to improve both nanoscale ordering and biosensing capabilities.

中文翻译:

低分子量蛋白质-聚合物通过蛋白质嵌段的低聚结合改善了有序排列

研究了掺入工程化的小亲和力结合蛋白rcSso7d.SA的寡聚物的蛋白质-聚合物共轭物的自组装,以确定蛋白质寡聚化对纳米级有序化的影响。寡聚化可在不改变其整体折叠结构的情况下系统地增加蛋白质摩尔质量,从而导致更高的驱动力,以自组装为有序结构。尽管发现单体rcSso7d.SA的缀合物仅以无序状态存在,但该蛋白的寡聚体与poly(N-异丙基丙烯酰胺(PNIPAM)块自组装成片状纳米结构。观察到三聚体和四聚体rcSso7d.SA的结合物在浓缩溶液中产生最强的有序性,在低至40 wt%的浓度下显示出双折射片。在高度浓缩的溶液中,低聚rcSso7d.SA-PNIPAM嵌段共聚物表现出与大多数嵌段共聚物不同的有序排列和畴间距变化趋势。荧光结合试验表明,寡聚蛋白块保留了结合功能,并显示出比表面固定蛋白传感器低三倍的检测限。因此,这些嵌段共聚物中蛋白质嵌段的低聚作用是提高纳米级有序性和生物传感能力的有效方法。
更新日期:2018-08-22
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