Using split protein reassembly strategy to optically control PLD enzymatic activity

yao, y.; Lou, X.; Li, J.; Zhu, M.; Qian, X.; Liu, J.; Zhang, L.; Zhang, P.; He, L.; Li, H.; Xu, Y.

doi:10.1101/2024.01.27.577557

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bioRxiv · 10.1101/2024.01.27.577557

Using split protein reassembly strategy to optically control PLD enzymatic activity

yao, y.·Lou, X.·Li, J.·Zhu, M.·Qian, X.·Liu, J.·Zhang, L.·Zhang, P.·He, L.·Li, H.·Xu, Y.

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Abstract

Phospholipase D (PLD) and phosphatidic acid (PA) play a spatio-temporal role in regulating diverse cellular activities. Although current methodologies enable optical control of the subcellular localization of PLD and by which influence local PLD enzyme activity, the overexpression of PLD elevates the basal PLD enzyme activity and further leads to increased PA levels in cells. In this study, we employed a split protein reassembly strategy and optogenetic techniques to modify superPLD (developed by Jeremy Baskin group). We splited this variants into two HKD domains and fused these domains with optogenetic and chemogenetic elements and by which we achieved control of the two HKD interaction and then restored the PLD enzymatic activity.

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yao, y., Lou, X., Li, J., Zhu, M., Qian, X., Liu, J., Zhang, L., Zhang, P., He, L., Li, H., Xu, Y.. 2024-01-30. Using split protein reassembly strategy to optically control PLD enzymatic activity. https://doi.org/10.1101/2024.01.27.577557

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