UPR 5301

Extending Janus lectins architecture: Characterization and application to protocells


“Synthetic biology is a rapidly growing field with applications in biotechnology and biomedicine. Through various approaches, remarkable achievements, such as cell and tissue engineering, have been already accomplished. In synthetic glycobiology, the engineering of glycan binding proteins is being exploited for producing tools with precise topology and specificity. We developed the concept of engineered chimeric lectins, i.e., Janus lectin, with increased valency, and additional specificity. The novel engineered lectin, assembled as a fusion protein between the b-propeller domain from Ralstonia solanacearum and the b-trefoil domain from fungus Marasmius oreades, is specific for fucose and a-galactose and its unique protein architecture allows to bind these ligands simultaneously. The protein activity was tested with glycosylated giant unilamellar vesicles, resulting in the formation of proto-tissue-like structures through cross-linking of such protocells. The engineered protein recognizes and binds H1299 human lung epithelial cancer cells by its two domains. The biophysical properties of this new construct were compared with the two already existing Janus lectins, RSL-CBM40 and RSL-CBM77 Rf. Denaturation profiles of the proteins indicate that the fold of each has a significant role in protein stability and should be considered during protein engineering. ”

The publication is available in open access.

The first author, Simona Nôtová, completed her thesis in our laboratory (2022), entitled “Engineering of Neo-Lctins and Lectins Janus”.
Summary: ️https: //cermav-test-bdd.local/en/blog/actualite/simona-notova-thesis-defense-on-june-29th-2022/

Simona was part of the Synbiocarb Innovative Training Network granted by the Research and Innovation Program in Europe “Marie Skłodowska-Curie Actions” (MSCA).

Such collaboration is also part of the “Innogly – Cost Action” project, the objective of which is to better understand the biological function of glycans in different biological contexts.