8ctp

Publication Abstract from PubMed

Cellular membranes contain numerous lipid species, and efforts to understand the biological functions of individual lipids have been stymied by a lack of approaches for controlled modulation of membrane composition in situ. Here we present a strategy for editing phospholipids, the most abundant lipids in biological membranes. Our membrane editor is based on a bacterial phospholipase D (PLD), which exchanges phospholipid head groups through hydrolysis or transphosphatidylation of phosphatidylcholine with water or exogenous alcohols. Exploiting activity-dependent directed enzyme evolution in mammalian cells, we have developed and structurally characterized a family of 'superPLDs' with up to a 100-fold enhancement in intracellular activity. We demonstrate the utility of superPLDs for both optogenetics-enabled editing of phospholipids within specific organelle membranes in live cells and biocatalytic synthesis of natural and unnatural designer phospholipids in vitro. Beyond the superPLDs, activity-based directed enzyme evolution in mammalian cells is a generalizable approach to engineer additional chemoenzymatic biomolecule editors.

Activity-based directed evolution of a membrane editor in mammalian cells.,Tei R, Bagde SR, Fromme JC, Baskin JM Nat Chem. 2023 Jul;15(7):1030-1039. doi: 10.1038/s41557-023-01214-0. Epub 2023 , May 22. PMID:37217787^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.