Membrane protein solubilization and structure determination using de novo–designed proteins
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.
Authors
- Edin Muratspahić (ORCID: https://orcid.org/0000-0002-6011-0805)
- Melissa J. Caimano (ORCID: https://orcid.org/0000-0003-1170-4102)
- Connor Weidle (ORCID: https://orcid.org/0000-0002-9354-2276)
- Alexis Courbet (ORCID: https://orcid.org/0000-0003-0539-7011)
- Neil P. King (ORCID: https://orcid.org/0000-0002-2978-4692)
- Ljubica Mihaljević (ORCID: https://orcid.org/0000-0002-3697-7767)
- Nicholas R. Thomson (ORCID: https://orcid.org/0000-0002-4432-8505)
- Helen E. Eisenach (ORCID: https://orcid.org/0000-0002-3201-4517)
- Rashmi Ravichandran (ORCID: https://orcid.org/0000-0002-2630-7487)
- Analisa Murray (ORCID: https://orcid.org/0000-0003-1560-6673)
- Thomas Schlichthaerle (ORCID: https://orcid.org/0000-0003-4444-2575)
- Andrew J. Borst (ORCID: https://orcid.org/0000-0003-4297-7824)
- Lorenzo Giacani (ORCID: https://orcid.org/0000-0003-3889-7274)
- Mila Lamb (ORCID: https://orcid.org/0000-0002-7318-1805)
- Linda Grillová (ORCID: https://orcid.org/0000-0003-2371-3161)
- David A. Baker (ORCID: https://orcid.org/0000-0001-7896-6217)
- David E. Kim (ORCID: https://orcid.org/0000-0002-0023-956X)
- Aldo T. Trejos (ORCID: https://orcid.org/0000-0002-7646-9839)
- Xinting Li (ORCID: https://orcid.org/0000-0001-5559-7578)
- Marc Expòsit (ORCID: https://orcid.org/0000-0002-2980-303X)
- Pengxiang Chang (ORCID: https://orcid.org/0000-0001-7000-2982)
- Everton B. Bettin (ORCID: https://orcid.org/0000-0001-5095-2198)
- Kelly L. Hawley (ORCID: https://orcid.org/0000-0001-8587-1662)
- Pooja Bandawane (ORCID: https://orcid.org/0000-0002-7729-5244)
- Kenneth D. Carr (ORCID: https://orcid.org/0009-0005-1929-3952)
- Qiushi Liu (ORCID: https://orcid.org/0009-0006-7992-0708)
- Sagardip Majumder (ORCID: https://orcid.org/0000-0002-4736-6945)
- Michael Landreh (ORCID: https://orcid.org/0000-0002-7958-4074)
- Surabhi Kokane
- Lynda Stuart
- Shuyuan Hu
- Alexander Stevens (ORCID: https://orcid.org/0009-0004-0773-0281)
- Elizabeth C. Williams
Institutions
- Uppsala University (SE)
- Centre National de la Recherche Scientifique (FR)
- Connecticut Department of Children and Families (US)
- Howard Hughes Medical Institute (US)
- Inserm (FR)
- University of Washington (US)
- Wellcome Sanger Institute (GB)
- Karolinska Institutet (SE)
- London School of Hygiene & Tropical Medicine (GB)
- Centre de Biologie Structurale (FR)
- Connecticut Children's Medical Center (US)
- Kymab (United Kingdom) (GB)
- Sanofi (United Kingdom) (GB)
- UConn Health (US)
Publication Details
- Journal
- Science
- Published
- 2026-07-02
- DOI
- https://doi.org/10.1126/science.adr3817
- Primary Topic
- Bacteriophages and microbial interactions
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Howard Hughes Medical Institute
- Bill and Melinda Gates Foundation
- National Institute of Allergy and Infectious Diseases