大连理工大学薛闯团队报道了使用刺激反应蛋白笼的动态代谢。相关研究成果于2024年3月1日发表在《美国化学会杂志》。

自然进化的代谢产物能够响应环境刺激进行组装和分解，从而能够快速重组活细胞中的化学反应，以满足不断变化的细胞需求。然而，由于人们对控制这种天然存在的多酶复合物的组装和拆卸的机制了解有限，在合成代谢产物中复制这种能力仍然是一个挑战。

该文中，研究人员报道了化学和光反应蛋白笼的合成，用于组装合成代谢产物，从而能够动态调节活细胞中的酶促反应。特别地，将化学反应结构域与自组装蛋白笼亚基融合，能够在其表面产生显示含有同源相互作用结构域的蛋白质的工程蛋白笼，响应小分子线索。化学诱导的序列酶在蛋白质笼上的共定位，将支链脱氧紫色杆菌素生物合成反应的特异性提高了2.6倍。

此外，通过用光诱导的二聚化结构域取代化学诱导结构域，研究人员创建了一个光遗传学蛋白质笼，该笼能够通过蓝光的开启和关闭在数十秒内，将靶向蛋白质可逆地募集到蛋白质笼的外部并从其释放。将光遗传学蛋白笼固定在膜上，可以形成可切换的、膜结合的代谢产物，这些代谢产物可以重复地聚集-释放酶，从而根据需要操纵跨膜的底物利用率。

该工作展示了一种在工程活细胞中构建动态代谢产物，以实现高效可控的生物催化的强大而通用的策略。

附：英文原文

Title: Dynamic Metabolons Using Stimuli-Responsive Protein Cages

Author: Wei Kang, Xiao Ma, Huawei Zhang, Juncai Ma, Chunxue Liu, Jiani Li, Hanhan Guo, Daping Wang, Rui Wang, Bo Li, Chuang Xue

Issue&Volume: March 1, 2024

Abstract: Naturally evolved metabolons have the ability to assemble and disassemble in response to environmental stimuli, allowing for the rapid reorganization of chemical reactions in living cells to meet changing cellular needs. However, replicating such capability in synthetic metabolons remains a challenge due to our limited understanding of the mechanisms by which the assembly and disassembly of such naturally occurring multienzyme complexes are controlled. Here, we report the synthesis of chemical- and light-responsive protein cages for assembling synthetic metabolons, enabling the dynamic regulation of enzymatic reactions in living cells. Particularly, a chemically responsive domain was fused to a self-assembled protein cage subunit, generating engineered protein cages capable of displaying proteins containing cognate interaction domains on their surfaces in response to small molecular cues. Chemical-induced colocalization of sequential enzymes on protein cages enhances the specificity of the branched deoxyviolacein biosynthetic reactions by 2.6-fold. Further, by replacing the chemical-inducible domain with a light-inducible dimerization domain, we created an optogenetic protein cage capable of reversibly recruiting and releasing targeted proteins onto and from the exterior of the protein cages in tens of seconds by on–off of blue light. Tethering the optogenetic protein cages to membranes enables the formation of light-switchable, membrane-bound metabolons, which can repeatably recruit–release enzymes, leading to the manipulation of substrate utilization across membranes on demand. Our work demonstrates a powerful and versatile strategy for constructing dynamic metabolons in engineered living cells for efficient and controllable biocatalysis.

DOI: 10.1021/jacs.3c12876

Source: https://pubs.acs.org/doi/abs/10.1021/jacs.3c12876