Enzymatic catalysis is essential to cell survival. In many instances, enzymes that participate in reaction cascades have been shown to assemble into metabolons in response to the presence of the substrate for the first enzyme. However, what triggers metabolon formation has remained an open question. Through a combination of theory and experiments, we show that enzymes in a cascade can assemble via chemotaxis. We apply microfluidic and fluorescent spectroscopy techniques to study the coordinated movement of the first four enzymes of the glycolysis cascade: hexokinase, phosphoglucose isomerase, phosphofructokinase and aldolase. We show that each enzyme independently follows its own specific substrate gradient, which in turn is produced by the preceding enzymatic reaction. Furthermore, we find that the chemotactic assembly of enzymes occurs even under cytosolic crowding conditions.
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The work was supported by Penn State MRSEC, funded by the National Science Foundation (NSF, DMR-1420620). P.J.B. acknowledges support from NSF CMMI 1334847. H.P. and H.H. acknowledge support by the Defense Threat Reduction Agency (award no. HDTRA1-14-1-0051).
M.K.G. has an equity interest in and is a cofounder and scientific advisor of VeraChem LLC.
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Zhao, X., Palacci, H., Yadav, V. et al. Substrate-driven chemotactic assembly in an enzyme cascade. Nature Chem 10, 311–317 (2018). https://doi.org/10.1038/nchem.2905
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