美国康奈尔大学教授Baskin, Jeremy M.团队报道了哺乳动物细胞膜编辑器基于活性的定向转化。相关研究成果于2023年5月22日发表在《自然—化学》。

细胞膜含有许多脂质物质，由于缺乏原位控制膜组成的方法，理解单个脂质的生物功能的努力受到阻碍。

该文中，研究人员提出了一种编辑生物膜中最丰富的脂质，磷脂的策略。该膜编辑器基于细菌磷脂酶D（PLD），它通过磷脂酰胆碱与水或外源醇的水解或转磷脂酰化来交换磷脂头基。利用哺乳动物细胞中活性依赖性的定向酶进化，研究人员开发了一个“超级PLD”家族，并对其进行了结构表征，该家族的细胞内活性提高了100倍。研究人员证明了超级PLD在活细胞中特定细胞器膜内磷脂的光遗传学编辑和体外天然和非天然设计磷脂的生物催化合成方面的实用性。除了超PLD之外，哺乳动物细胞中基于活性的定向酶进化是一种通用的方法，可以设计额外的化学酶生物分子编辑器。

附：英文原文

Title: Activity-based directed evolution of a membrane editor in mammalian cells

Author: Tei, Reika, Bagde, Saket R., Fromme, J. Christopher, Baskin, Jeremy M.

Issue&Volume: 2023-05-22

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.

DOI: 10.1038/s41557-023-01214-0

Source: https://www.nature.com/articles/s41557-023-01214-0