8JGC

Cryo-EM structure of Mi3 fused with LOV2

Summary for 8JGC

• Entry DOI: 10.2210/pdb8jgc/pdb
• EMDB information: 36230
• Descriptor: LOV domain-containing protein,2-dehydro-3-deoxyphosphogluconate aldolase/4-hydroxy-2-oxoglutarate aldolase (1 entity in total)
• Functional Keywords: protein cage scaffold, lyase
• Biological source: Aegilops tauschii subsp. strangulata (Goatgrass); More
• Total number of polymer chains: 1
• Total formula weight: 41424.47

Authors

Zhang, H.W.,Kang, W.,Xue, C. (deposition date: 2023-05-20, release date: 2024-04-24, Last modification date: 2024-10-23)

Primary citation

Kang, W.,Ma, X.,Zhang, H.,Ma, J.,Liu, C.,Li, J.,Guo, H.,Wang, D.,Wang, R.,Li, B.,Xue, C.
Dynamic Metabolons Using Stimuli-Responsive Protein Cages.
J.Am.Chem.Soc., 146:6686-6696, 2024

PubMed 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.

PubMed: 38425051
DOI: 10.1021/jacs.3c12876

Experimental method

ELECTRON MICROSCOPY (3.44 Å)