8CTQ

Crystal structure of engineered phospholipase D mutant superPLD 2-48

8CTQ の概要

• エントリーDOI: 10.2210/pdb8ctq/pdb
• 関連するPDBエントリー: 8CTP
• 分子名称: Phospholipase D, PHOSPHATE ION (3 entities in total)
• 機能のキーワード: enzyme, phospholipase, pld, phospholipid, hydrolase
• 由来する生物種: Streptomyces sp. PMF
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 54891.14

構造登録者

Tei, R.,Bagde, S.R.,Fromme, J.C.,Baskin, J.M. (登録日: 2022-05-16, 公開日: 2023-04-26, 最終更新日: 2024-11-13)

主引用文献

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

実験手法

X-RAY DIFFRACTION (1.85 Å)