Rapid Formation of Non‐canonical Phospholipid Membranes by Chemoselective Amide‐Forming Ligations with Hydroxylamines

• Published in: Angewandte Chemie 2024-01, Vol.136 (1), p.n/a
• Main Authors: Chen, Jiyue, Brea, Roberto J., Fracassi, Alessandro, Cho, Christy J., Wong, Adrian M., Salvador‐Castell, Marta, Sinha, Sunil K., Budin, Itay, Devaraj, Neal K.
• Format: Article
• Language: English
• Subjects: Biocompatibility; Bioconjugation; Biosynthesis; Cells (biology); Chemical reactions; Chemistry; Hydroxylamines; Lipid membranes; Lipids; Membranes; Phospholipids; Physiology; Precursors; Self-Assembly
• ISSN: 0044-8249; 1521-3757
• DOI: 10.1002/ange.202311635

There has been increasing interest in methods to generate synthetic lipid membranes as key constituents of artificial cells or to develop new tools for remodeling membranes in living cells. However, the biosynthesis of phospholipids involves elaborate enzymatic pathways that are challenging to reconstitute in vitro. An alternative approach is to use chemical reactions to non‐enzymatically generate natural or non‐canonical phospholipids de novo. Previous reports have shown that synthetic lipid membranes can be formed in situ using various ligation chemistries, but these methods lack biocompatibility and/or suffer from slow kinetics at physiological pH. Thus, it would be valuable to develop chemoselective strategies for synthesizing phospholipids from water‐soluble precursors that are compatible with synthetic or living cells Here, we demonstrate that amide‐forming ligations between lipid precursors bearing hydroxylamines and α‐ketoacids (KAs) or potassium acyltrifluoroborates (KATs) can be used to prepare non‐canonical phospholipids at physiological pH conditions. The generated amide‐linked phospholipids spontaneously self‐assemble into cell‐like micron‐sized vesicles similar to natural phospholipid membranes. We show that lipid synthesis using KAT ligation proceeds extremely rapidly, and the high selectivity and biocompatibility of the approach facilitates the in situ synthesis of phospholipids and associated membranes in living cells. Bioorthogonal de novo phospholipid synthesis utilizing the reactions between hydroxylamine and α‐ketoacids (KAs) or potassium acyltrifluoroborates (KATs) enables the rapid formation of biomimetic membranes under physiological pH conditions, using reactant concentrations in the micromolar range. Additionally, these reactions are biocompatible and can be performed in living mammalian cells.