Moinpour Mahta, Fracassi Alessandro, Brea Roberto J, Salvador-Castell Marta, Pandey Sudip, Edwards Madison M, Seifert Soenke, Joseph Simpson, Sinha Sunil K, Devaraj Neal K
Department of Chemistry and Biochemistry, University of California, San Diego, 9500 Gilman Drive, Natural Sciences Building 3328, La Jolla, CA 92093, USA.
Biomimetic Membrane Chemistry (BioMemChem) Group, Centro de Investigacións Científicas Avanzadas (CICA), Universidade da Coruña, Rúa As Carballeiras, 15701, A Coruña, Spain.
Chembiochem. 2022 Mar 4;23(5):e202100624. doi: 10.1002/cbic.202100624. Epub 2022 Jan 19.
All cells use organized lipid compartments to facilitate specific biological functions. Membrane-bound organelles create defined spatial environments that favor unique chemical reactions while isolating incompatible biological processes. Despite the fundamental role of cellular organelles, there is a scarcity of methods for preparing functional artificial lipid-based compartments. Here, we demonstrate a robust bioconjugation system for sequestering proteins into zwitterionic lipid sponge phase droplets. Incorporation of benzylguanine (BG)-modified phospholipids that form stable covalent linkages with an O -methylguanine DNA methyltransferase (SNAP-tag) fusion protein enables programmable control of protein capture. We show that this methodology can be used to anchor hydrophilic proteins at the lipid-aqueous interface, concentrating them within an accessible but protected chemical environment. SNAP-tag technology enables the integration of proteins that regulate complex biological functions in lipid sponge phase droplets, and should facilitate the development of advanced lipid-based artificial organelles.
所有细胞都利用有组织的脂质区室来促进特定的生物学功能。膜结合细胞器创造了明确的空间环境，有利于独特的化学反应，同时隔离不相容的生物学过程。尽管细胞器具有重要作用，但制备功能性人工脂质区室的方法却很匮乏。在这里，我们展示了一种强大的生物共轭系统，用于将蛋白质隔离到两性离子脂质海绵相液滴中。掺入与O-甲基鸟嘌呤DNA甲基转移酶（SNAP标签）融合蛋白形成稳定共价键的苄基鸟嘌呤（BG）修饰磷脂，能够对蛋白质捕获进行可编程控制。我们表明，这种方法可用于将亲水蛋白质锚定在脂质-水界面，将它们集中在一个可及但受保护的化学环境中。SNAP标签技术能够将调节复杂生物学功能的蛋白质整合到脂质海绵相液滴中，应该会促进先进的基于脂质的人工细胞器的发展。
