Controlling Protein Enrichment in Lipid Sponge Phase Droplets using SNAP–Tag Bioconjugation

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.

doi:10.1002/cbic.202100624

Controlling Protein Enrichment in Lipid Sponge Phase Droplets using SNAP–Tag Bioconjugation

Journal Article · Wed Jan 19 00:00:00 EST 2022 · ChemBioChem: a European journal of chemical biology

DOI:https://doi.org/10.1002/cbic.202100624· OSTI ID:1879855

Moinpour, Mahta ^[1]; ^[1]; ^[2]; ^[1]; ^[1]; ^[1]; ^[3]; ^[1]; ^[1]; ^[1]

Univ. of California, San Diego, La Jolla, CA (United States)
Universidade da Coruña (Spain)
Argonne National Lab. (ANL), Lemont, IL (United States)

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. In this work, 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.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: US Army Research Office (ARO); USDOE Office of Science (SC), Basic Energy Sciences (BES); University of California; Xunta de Galicia

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1879855

Journal Information:: ChemBioChem: a European journal of chemical biology, Journal Name: ChemBioChem: a European journal of chemical biology Journal Issue: 5 Vol. 23; ISSN 1439-4227

Publisher:: ChemPubSoc EuropeCopyright Statement

Country of Publication:: United States

Language:: English

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