Dynamic Metabolons Using Stimuli-Responsive Protein Cages

作者：Wei Kang, Xiao Ma, Huawei Zhang, Juncai Ma, Chunxue Liu, Jiani Li, Hanhan Guo, Daping Wang, Rui Wang, Bo Li, Chuang Xue · 发表于：Journal of the American Chemical Society · 年份：2024 · DOI：10.1021/jacs.3c12876 · 被引用次数：21 · 研究领域：Bacteriophages and microbial interactions、Monoclonal and Polyclonal Antibodies Research、Glycosylation and Glycoproteins Research

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...