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Recently, photo\u2010sensitive lipids were introduced to study membrane domain organization, and photo\u2010isomerization was shown to trigger the mixing and de\u2010mixing of liquid\u2010ordered (<jats:italic>l<\/jats:italic><jats:sub>o<\/jats:sub>) domains in artificial phase\u2010separated membranes. Here, we synthesized globotriaosylceramide (Gb<jats:sub>3<\/jats:sub>) glycosphingolipids that harbor an azobenzene moiety at different positions of the fatty acid to investigate light\u2010induced membrane domain reorganization, and that serve as specific receptors for the protein Shiga toxin (STx). Using phase\u2010separated supported lipid bilayers on mica surfaces doped with four different photo\u2010Gb<jats:sub>3<\/jats:sub> molecules, we found by fluorescence microscopy and atomic force microscopy that liquid disordered (<jats:italic>l<\/jats:italic><jats:sub>d<\/jats:sub>) domains were formed within <jats:italic>l<\/jats:italic><jats:sub>o<\/jats:sub> domains upon <jats:italic>trans<\/jats:italic>\u2010<jats:italic>cis<\/jats:italic> photo\u2010isomerization. The fraction and size of these <jats:italic>l<\/jats:italic><jats:sub>d<\/jats:sub> domains were largest for Gb<jats:sub>3<\/jats:sub> molecules with the azobenzene group at the end of the fatty acid. We further investigated the impact of domain reorganization on the interaction of the B\u2010subunits of STx with the photo\u2010Gb<jats:sub>3<\/jats:sub>. Fluorescence and atomic force micrographs clearly demonstrated that STxB binds to the <jats:italic>l<\/jats:italic><jats:sub>o<\/jats:sub> phase if Gb<jats:sub>3<\/jats:sub> is in the <jats:italic>trans<\/jats:italic>\u2010configuration, whereas two STxB populations are formed if the photo\u2010Gb<jats:sub>3<\/jats:sub> is switched to the <jats:italic>cis<\/jats:italic>\u2010configuration highlighting the idea of manipulating lipid\u2010protein interactions with a light stimulus.<\/jats:p>","DOI":"10.1002\/chem.202202766","type":"journal-article","created":{"date-parts":[[2022,10,24]],"date-time":"2022-10-24T17:56:45Z","timestamp":1666634205000},"update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Optical Manipulation of Gb<sub>3<\/sub> Enriched Lipid Domains: Impact of Isomerization on Gb<sub>3<\/sub>\u2010Shiga Toxin B 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