Tunable Extracellular Self-Assembly of Multi-Protein Conjugates from Bacillus subtilis
Tunable Extracellular Self-Assembly of Multi-Protein Conjugates from Bacillus subtilis
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DOI：
10.1101/087593
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发表时间：
2016-11
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通讯作者：
Charlie Gilbert;M. Howarth;C. Harwood;T. Ellis
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作者：
Charlie Gilbert;M. Howarth;C. Harwood;T. Ellis
重组蛋白彼此稳定、特异性结合的能力是一种强大的体外技术，可用于工程多功能酶、蛋白治疗剂和新型生物材料。然而，对于许多应用来说，自发的体内蛋白质缀合比体外方法更可取。利用最近描述的 SpyTag-SpyCatcher 系统，我们在此描述了如何对酶和结构蛋白进行基因编码，以在枯草芽孢杆菌可编程分泌后在培养基中共价结合。使用这种新方法，我们证明了分泌型工业酶 XynA 的自结合如何显着提高其对沸腾的适应能力，并且我们表明细胞联合体可以被设计为自组装具有可调组成的功能性多蛋白复合物。这种基因编码的模块化系统利用细胞外自组装的巨大优势，为蛋白质缀合提供了一种新的、灵活的策略。
The ability to stably and specifically conjugate recombinant proteins to one another is a powerful in vitro technique for engineering multifunctional enzymes, protein therapeutics and novel biological materials. However, for many applications spontaneous in vivo protein conjugation would be preferable to in vitro methods. Exploiting the recently described SpyTag-SpyCatcher system, we describe here how enzymes and structural proteins can be genetically-encoded to covalently conjugate in culture media following programmable secretion by Bacillus subtilis. Using this novel approach, we demonstrate how self-conjugation of a secreted industrial enzyme, XynA, dramatically increases its resilience to boiling and we show that cellular consortia can be engineered to self-assemble functional multi-protein complexes with tunable composition. This genetically-encoded modular system provides a new, flexible strategy for protein conjugation harnessing the substantial advantages of extracellular self-assembly.