Peer-reviewedCC BYScience (New York, N.Y.)
Membrane protein solubilization and structure determination using de novo–designed proteins
6 affiliations
- 1Department of Biochemistry, Institute for Protein Design, University of Washington, Seattle, WA 98195, USA.
- 2Department of Pediatrics, UConn Health, Farmington, CT 06030-3715, USA.
- 3Kymab, a Sanofi Company, Babraham Research Campus, Cambridge, UK.
- 4Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington
- 5Department of Cell and Molecular Biology, Uppsala University, 751 24 Uppsala, Sweden
- 6Parasites and Microbes Programme, Wellcome Sanger Institute, Hinxton, UK.
Abstract
Developing therapies and vaccines against integral membrane proteins is hindered by their extensive hydrophobic surfaces, which complicate production and structural analysis. Here, we describe a general deep learning–based design approach for solubilizing native membrane proteins while preserving their sequence, fold, active-site, and ligand-binding properties. Genetically encoded de novo protein WRAPs [water-soluble RFdiffused amphipathic proteins] surround the lipid-interacting hydrophobic surfaces, rendering them thermostable and water-soluble without the need for detergents. We design WRAPs for both monomeric and oligomeric beta-barrel outer membrane proteins and helical multipass transmembrane proteins. A 2.95-angstrom-resolution cryo–electron microscopy structure of WRAPed mycobacterial porin demonstrates that WRAPs can be used for the structural determination of membrane proteins in solution. As a step toward syphilis vaccine development, we generated soluble versions of Treponema pallidum antigens.
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BibTeX
@article{mihaljevic2026membrane,
title = {{Membrane protein solubilization and structure determination using de novo–designed proteins}},
author = {Ljubica Mihaljević and David E. Kim and Pooja D. Bandawane and Helen E. Eisenach and Andrew J. Borst and Alexis Courbet and Connor Weidle and Kenneth D. Carr and Everton Bettin and Qiushi Liu and Aldo T. Trejos and Sagardip Majumder and Surabhi Kokane and Alexander Stevens and Edin Muratspahić and Thomas Schlichthaerle and Marc Expòsit and Xinting Li and Mila Lamb and Analisa Nicole Azcárraga Murray and Rashmi Ravichandran and Elizabeth C. Williams and Shuyuan Hu and Lynda Stuart and Linda Grillová and Nicholas R. Thomson and Michael Landreh and Pengxiang Chang and Lorenzo Giacani and Melissa J. Caimano and Kelly L. Hawley and Neil P. King and David Baker},
journal = {Science (New York, N.Y.)},
year = {2026},
month = jul,
doi = {10.1126/science.adr3817},
url = {https://europepmc.org/article/PMC/PMC13730216},
note = {CC BY},
}
RIS
TY - JOUR
TI - Membrane protein solubilization and structure determination using de novo–designed proteins
AU - Ljubica Mihaljević
AU - David E. Kim
AU - Pooja D. Bandawane
AU - Helen E. Eisenach
AU - Andrew J. Borst
AU - Alexis Courbet
AU - Connor Weidle
AU - Kenneth D. Carr
AU - Everton Bettin
AU - Qiushi Liu
AU - Aldo T. Trejos
AU - Sagardip Majumder
AU - Surabhi Kokane
AU - Alexander Stevens
AU - Edin Muratspahić
AU - Thomas Schlichthaerle
AU - Marc Expòsit
AU - Xinting Li
AU - Mila Lamb
AU - Analisa Nicole Azcárraga Murray
AU - Rashmi Ravichandran
AU - Elizabeth C. Williams
AU - Shuyuan Hu
AU - Lynda Stuart
AU - Linda Grillová
AU - Nicholas R. Thomson
AU - Michael Landreh
AU - Pengxiang Chang
AU - Lorenzo Giacani
AU - Melissa J. Caimano
AU - Kelly L. Hawley
AU - Neil P. King
AU - David Baker
PY - 2026
DA - 2026/07/02
JO - Science (New York, N.Y.)
DO - 10.1126/science.adr3817
UR - https://europepmc.org/article/PMC/PMC13730216
AB - Developing therapies and vaccines against integral membrane proteins is hindered by their extensive hydrophobic surfaces, which complicate production and structural analysis. Here, we describe a general deep learning–based design approach for solubilizing native membrane proteins while preserving their sequence, fold, active-site, and ligand-binding properties. Genetically encoded de novo protein WRAPs [water-soluble RFdiffused amphipathic proteins] surround the lipid-interacting hydrophobic surfaces, rendering them thermostable and water-soluble without the need for detergents. We design WRAPs for both monomeric and oligomeric beta-barrel outer membrane proteins and helical multipass transmembrane proteins. A 2.95-angstrom-resolution cryo–electron microscopy structure of WRAPed mycobacterial porin demonstrates that WRAPs can be used for the structural determination of membrane proteins in solution. As a step toward syphilis vaccine development, we generated soluble versions of Treponema pallidum antigens.
SN - 0036-8075
SN - 1095-9203
ER -