Rational design of elastin-like polypeptide fusion proteins to tune self-assembly and properties of protein vesicles
Rational design of elastin-like polypeptide fusion proteins to tune self-assembly and properties of protein vesicles
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DOI：
10.1039/d3tb00200d
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发表时间：
2023-06-26
影响因子：
7
通讯作者：
Champion, Julie A.
中科院分区：
文献类型：
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作者：
Li, Yirui;Dautel, Dylan R.;Champion, Julie A.
由生物活性蛋白制成的蛋白囊泡在药物传递、生物催化和人造细胞方面具有潜在的价值。由于蛋白质是重组产生的，精确调整蛋白质序列的能力提供了聚合物囊泡无法实现的控制。蛋白质的可调节性和生物相容性促使本研究利用合理设计的蛋白质构建块来开发蛋白质囊泡，以研究蛋白质序列如何影响囊泡的自组装和性质。我们报道了一种弹性蛋白样多肽（ELP）融合到富含精氨酸的亮氨酸拉链上（Z(R)），以及一种功能性球状蛋白融合到富含谷氨酸的亮氨酸拉链上（Z(E)），当温度从4℃升高到25℃时，它们会自组装成蛋白质囊泡；C是由于ELP的疏水转变。之前，我们证明了通过改变蛋白质和盐浓度、Z(E): Z(R)比和升温速率来调节囊泡特性的能力。但是，通过装配条件可以实现的属性是有限的。为了获得更大范围的囊泡直径和稳定性，本研究以mCherry为模型球状蛋白，研究了修饰ELP序列的疏水性和长度如何影响蛋白囊泡的自组装和最终特性。结果表明，转变温度和蛋白囊泡直径与ELP客体残基疏水性和ELP五肽重复数呈负相关。此外，序列操作允许装配具有不能通过更改装配条件访问的属性的囊泡。例如，在ELP的5个客体残基位置引入酪氨酸，可以形成在生理盐浓度下稳定的纳米级囊泡。这项工作产生了修改ELP序列的设计指南，以操纵蛋白质囊泡转变温度、大小和稳定性，以实现特定生物功能应用所需的特性。
Protein vesicles made from bioactive proteins have potential value in drug delivery, biocatalysis, and as artificial cells. As the proteins are produced recombinantly, the ability to precisely tune the protein sequence provides control not possible with polymeric vesicles. The tunability and biocompatibility motivated this work to develop protein vesicles using rationally designed protein building blocks to investigate how protein sequence influences vesicle self-assembly and properties. We have reported an elastin-like polypeptide (ELP) fused to an arginine-rich leucine zipper (Z(R)) and functional, globular proteins fused to a glutamate-rich leucine zipper (Z(E)) that self-assemble into protein vesicles when warmed from 4 to 25 & DEG;C due to the hydrophobic transition of ELP. Previously, we demonstrated the ability to tune vesicle properties by changing protein and salt concentration, Z(E) : Z(R) ratio, and warming rate. However, there is a limit to the properties that can be achieved via assembly conditions. In order to access a wider range of vesicle diameter and stability profiles, this work investigated how modifiying the hydrophobicity and length of the ELP sequence influenced self-assembly and the final properties of protein vesicles using mCherry as a model globular protein. The results showed that both transition temperature and diameter of protein vesicles were inversely correlated to the ELP guest residue hydrophobicity and the number of ELP pentapeptide repeats. Additionally, sequence manipulation enabled assembly of vesicles with properties not accessible by changes to assembly conditions. For example, introduction of tyrosine at 5 guest residue positions in ELP enabled formation of nanoscale vesicles stable at physiological salt concentration. This work yields design guidelines for modifying the ELP sequence to manipulate protein vesicle transition temperature, size and stability to achieve desired properties for particular biofunctional applications.