8CTP

Crystal structure of engineered phospholipase D mutant superPLD 2-23

Summary for 8CTP

• Entry DOI: 10.2210/pdb8ctp/pdb
• Descriptor: Phospholipase D, PHOSPHATE ION (3 entities in total)
• Functional Keywords: enzyme, phospholipase, pld, phospholipid, hydrolase
• Biological source: Streptomyces sp. PMF
• Total number of polymer chains: 1
• Total formula weight: 54811.07

Authors

Tei, R.,Bagde, S.R.,Fromme, J.C.,Baskin, J.M. (deposition date: 2022-05-16, release date: 2023-04-26, Last modification date: 2024-11-06)

Primary citation

Tei, R.,Bagde, S.R.,Fromme, J.C.,Baskin, J.M.
Activity-based directed evolution of a membrane editor in mammalian cells.
Nat.Chem., 15:1030-1039, 2023

PubMed Abstract: 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.

PubMed: 37217787
DOI: 10.1038/s41557-023-01214-0

Experimental method

X-RAY DIFFRACTION (1.9 Å)