Designed Artificial Protein Heterodimers With Coupled Functions Constructed Using Bio-Orthogonal Chemistry.
Designed Artificial Protein Heterodimers With Coupled Functions Constructed Using Bio-Orthogonal Chemistry.
复制标题
设计的人造蛋白质异二聚体具有使用生物正交化学构建的耦合功能。
DOI：
10.3389/fchem.2021.733550
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发表时间：
2021
影响因子：
5.5
通讯作者：
Jones DD
中科院分区：
文献类型：
--
作者：
Johnson RL;Blaber HG;Evans T;Worthy HL;Pope JR;Jones DD
蛋白质复合物的形成是生物学的核心，寡聚蛋白质比单体更普遍。功能上甚至结构上不同的蛋白质单元的偶联可以导致单体蛋白质单独无法获得的新功能特性。虽然这种复合物是由生物学的进化需求驱动的，但将功能和结构不同的蛋白质正常连接起来的能力可能会导致新的新兴特性用于合成生物学和纳米科学。在这里，我们展示了两个不同的蛋白质，血红素结合螺旋束蛋白细胞色素B 562和β-桶绿色荧光蛋白可以结合，形成一个异二聚体连接在一起的非天然三唑键。该复合物的设计使用计算对接方法来预测两种蛋白质之间的相容界面。然后，复合物的模型用于设计每个蛋白质中的残基偶联位点，以将它们连接在一起。遗传密码扩展用于在细胞色素B 562中引入叠氮化物化学和在GFP中引入炔化学，使得可以在两种蛋白质之间形成永久的三唑共价键。对GFP的两个连锁位点进行取样。新的异源二聚体的光谱分析表明，血红素结合和荧光蛋白的发色团的性质被保留。通过GFP吸光度和荧光的变化证实了功能偶联，其中连接位点决定了两种蛋白质之间的通信程度。因此，我们在这里已经表明，有可能设计和构建异二聚体蛋白质，这些蛋白质在结构和功能上不同，从而形成具有新功能特性的新复合物。
The formation of protein complexes is central to biology, with oligomeric proteins more prevalent than monomers. The coupling of functionally and even structurally distinct protein units can lead to new functional properties not accessible by monomeric proteins alone. While such complexes are driven by evolutionally needs in biology, the ability to link normally functionally and structurally disparate proteins can lead to new emergent properties for use in synthetic biology and the nanosciences. Here we demonstrate how two disparate proteins, the haem binding helical bundle protein cytochrome b 562 and the β-barrel green fluorescent protein can be combined to form a heterodimer linked together by an unnatural triazole linkage. The complex was designed using computational docking approaches to predict compatible interfaces between the two proteins. Models of the complexes where then used to engineer residue coupling sites in each protein to link them together. Genetic code expansion was used to incorporate azide chemistry in cytochrome b 562 and alkyne chemistry in GFP so that a permanent triazole covalent linkage can be made between the two proteins. Two linkage sites with respect to GFP were sampled. Spectral analysis of the new heterodimer revealed that haem binding and fluorescent protein chromophore properties were retained. Functional coupling was confirmed through changes in GFP absorbance and fluorescence, with linkage site determining the extent of communication between the two proteins. We have thus shown here that is possible to design and build heterodimeric proteins that couple structurally and functionally disparate proteins to form a new complex with new functional properties.