Controlling Protein Enrichment in Lipid Sponge Phase Droplets using SNAP‐Tag Bioconjugation
Abstract
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 O6‐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.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jan 19, 2022
- Source ID
- 10.1002/cbic.202100624
Entities
People
- Alessandro Fracassi
- M Salvador Castell
- Madison Edwards
- Mahta Moinpour
- Neal Devaraj
- Roberto Javier Brea
- S. K. Sinha
- Simpson Joseph
- Soenke Seifert
- Sudip Pandey
Organizations
- Argonne National Laboratory
- Stanford University
- University of California, San Diego
- Xunta de Galicia