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Color-coded galectin fusion proteins as novel tools in biomaterial science
Carina Dey1, Isabel K Sommerfeld2,3, Pavla Bojarová4,5
1Laboratory for Biomaterials, Institute for Biotechnology and Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University, Pauwelsstraße 20, 52074 Aachen, Germany. l.elling@biotec.rwth-aachen.de.
Researchers developed novel galectin fusion proteins for advanced biotechnological and biomedical uses. These engineered proteins enable precise molecular assembly and targeted delivery, showing promise in cancer cell applications.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Human galectins possess inherent carbohydrate-binding specificities valuable for biotechnological and biomedical applications.
- Fusion of galectin carbohydrate-recognition domains (CRDs) with peptides or proteins allows for multifunctional applications including purification, immobilization, and imaging.
Purpose of the Study:
- To create a library of color-coded galectin fusion proteins for versatile immobilization and imaging.
- To assess the impact of N- and C-terminal fusion partners on galectin binding properties and SpyCatcher/SpyTag immobilization efficacy.
- To demonstrate the utility of these fusion proteins in constructing extracellular matrix (ECM)-like structures and in targeted delivery applications.
Main Methods:
- Construction of a library of galectin fusion proteins incorporating His6-tag, fluorescent protein, and SpyCatcher/SpyTag units.
- Characterization of galectin fusion protein binding affinities and SpyCatcher/SpyTag-mediated immobilization.
- In vitro assembly of a three-layer ECM-like structure using SC-ST-Gal-3 conjugates.
- Immobilization of galectins in microgels for localized targeting applications.
Main Results:
- Galectin fusion proteins retained similar binding properties to non-fused galectins, with micromolar apparent binding affinities.
- N- and C-terminal fusion partners did not impede SpyCatcher/SpyTag immobilization.
- Successful stepwise formation of an ECM-like structure in vitro was achieved.
- Demonstrated galectin-mediated binding of microgels to colorectal cancer cells, validating their potential in targeted delivery.
Conclusions:
- The developed galectin fusion proteins offer a versatile platform for biotechnological and biomedical applications.
- SpyCatcher/SpyTag technology effectively enables galectin immobilization for constructing complex structures and targeted delivery systems.
- These engineered galectins show significant potential for localized targeting strategies, particularly in cancer therapy.
