Applications in biotechnology and synthetic biology often make use of soluble proteins, but there are many potential advantages to anchoring enzymes to a stable substrate, including stability and the possibility for substrate channeling. To avoid the necessity of protein purification and chemical immobilization, there has been growing interest in bio-assembly of protein-containing nanoparticles, exploiting the self-assembly of viral capsid proteins or other proteins that form polyhedral structures. But these nanoparticle are limited in size which constrains the packaging and the accessibility of the proteins. The axoneme, the insoluble protein core of the eukaryotic flagellum or cilium, is a highly ordered protein structure that can be several microns in length, orders of magnitude larger than other types of nanoparticles. We show that when proteins of interest are fused to specific axonemal proteins and expressed in living cells, they become incorporated into linear arrays which have the advantages of high protein loading capacity, high stability, and single-step purification with retention of biomass. The arrays can be isolated as membrane enclosed vesicle or as exposed protein arrays. The approach is demonstrated for both fluorescent proteins and enzymes, and in the latter case it is found that incorporation into axoneme arrays provides increased stability for the enzyme.

中文翻译：

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使用鞭毛轴丝生物合成线性蛋白质纳米阵列

生物技术和合成生物学中的应用通常使用可溶性蛋白质，但将酶锚定到稳定的底物上有许多潜在的优势，包括稳定性和底物通道的可能性。为了避免蛋白质纯化和化学固定的必要性，利用病毒衣壳蛋白或形成多面体结构的其他蛋白质的自组装，对含蛋白质纳米颗粒的生物组装越来越感兴趣。但是这些纳米颗粒的尺寸有限，这限制了蛋白质的包装和可及性。轴丝是真核生物鞭毛或纤毛的不溶性蛋白质核心，是一种高度有序的蛋白质结构，长度可达数微米，比其他类型的纳米粒子大几个数量级。我们表明，当感兴趣的蛋白质与特定的轴索蛋白融合并在活细胞中表达时，它们会被整合到线性阵列中，该阵列具有高蛋白质负载能力、高稳定性和可保留生物量的单步纯化等优点。阵列可以分离为膜封闭囊泡或暴露的蛋白质阵列。该方法适用于荧光蛋白和酶，在后一种情况下，发现掺入轴丝阵列为酶提供了更高的稳定性。阵列可以分离为膜封闭囊泡或暴露的蛋白质阵列。该方法适用于荧光蛋白和酶，在后一种情况下，发现掺入轴丝阵列为酶提供了更高的稳定性。阵列可以分离为膜封闭囊泡或暴露的蛋白质阵列。该方法适用于荧光蛋白和酶，在后一种情况下，发现掺入轴丝阵列为酶提供了更高的稳定性。