The effects of protein charge patterning on complex coacervation
The effects of protein charge patterning on complex coacervation
复制标题
蛋白质电荷模式对复合凝聚的影响
DOI：
10.1039/d1sm00543j
复制
发表时间：
2021
期刊：
影响因子：
3.4
通讯作者：
Obermeyer, Allie C.
中科院分区：
文献类型：
--
作者：
Zervoudis, Nicholas A.;Obermeyer, Allie C.
蛋白质与其他大分子的复合凝聚在蛋白质包封和递送以及用于确定细胞凝聚体的功能中具有应用。蛋白质凝聚体的理论或经验预测将使这些凝聚体的设计与可调和可预测的结构-功能关系，不幸的是，没有这样的理论存在。为了帮助建立预测模型，本研究探讨了蛋白质特异性参数对复凝聚的影响。序列特异性的、多肽标记的GFP变体和强合成酶的复合凝聚用于评估蛋白质电荷图案化对相行为的影响。蛋白质凝聚体的相图表明，电荷图案决定了蛋白质的双节点相边界。在凝聚相中达到超过100 mg mL-1的蛋白质浓度，浓度取决于共价连接到球状蛋白结构域的标签多肽序列。除了移动的双节相边界，多肽电荷图案化提供熵的优势，各向同性图案化的蛋白质。总之，这些结果表明，标签多肽序列中仅几个氨基酸的适度变化改变凝聚热力学，并可用于调节感兴趣的多肽或蛋白质的相行为。
The complex coacervation of proteins with other macromolecules has applications in protein encapsulation and delivery and for determining the function of cellular coacervates. Theoretical or empirical predictions for protein coacervates would enable the design of these coacervates with tunable and predictable structure–function relationships; unfortunately, no such theories exist. To help establish predictive models, the impact of protein-specific parameters on complex coacervation were probed in this study. The complex coacervation of sequence-specific, polypeptide-tagged, GFP variants and a strong synthetic polyelectrolyte was used to evaluate the effects of protein charge patterning on phase behavior. Phase portraits for the protein coacervates demonstrated that charge patterning dictates the protein's binodal phase boundary. Protein concentrations over 100 mg mL−1 were achieved in the coacervate phase, with concentrations dependent on the tag polypeptide sequence covalently attached to the globular protein domain. In addition to shifting the binodal phase boundary, polypeptide charge patterning provided entropic advantages over isotropically patterned proteins. Together, these results show that modest changes of only a few amino acids in the tag polypeptide sequence alter the coacervation thermodynamics and can be used to tune the phase behavior of polypeptides or proteins of interest.