Development of a Novel Cell Surface Attachment System to Display Multi-Protein Complex Using the Cohesin-Dockerin Binding Pair
Autor: | Hyeok-Jin Ko, Heesang Song, In Geol Choi |
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Rok vydání: | 2021 |
Předmět: |
Protein Folding
Multiprotein complex Chromosomal Proteins Non-Histone Archaeal Proteins Recombinant Fusion Proteins Cell Dockerin Cell Cycle Proteins medicine.disease_cause Disaccharidases Applied Microbiology and Biotechnology Green fluorescent protein Hydrolase medicine Escherichia coli Amino Acid Sequence Cohesin Chemistry Cell Membrane General Medicine Folding (chemistry) Luminescent Proteins medicine.anatomical_structure Archaeoglobus fulgidus Biophysics Cell Surface Display Techniques Biotechnology Protein Binding |
Zdroj: | Journal of microbiology and biotechnology. 31(8) |
ISSN: | 1738-8872 |
Popis: | Autodisplay of a multimeric protein complex on a cell surface is limited by intrinsic factors such as the types and orientations of anchor modules. Moreover, improper folding of proteins to be displayed often hinders functional cell surface display. While overcoming these drawbacks, we ultimately extended the applicability of the autodisplay platform to the display of a protein complex. We designed and constructed a cell surface attachment (CSA) system that uses a non-covalent protein-protein interaction. We employed the high-affinity interaction mediated by an orthogonal cohesin (Coh)-dockerin (Doc) pair from Archaeoglobus fulgidus to build the CSA system. Then, we validated the orthogonal binding between Coh and Doc by the attachment of monomeric red fluorescent protein to the cell surface. We evaluated the functional anchoring of proteins fused with the Doc module to the autodisplayed Coh module on the surface of Escherichia coli. The designed CSA system was applied to make a functional attachment of dimeric α-neoagarobiose hydrolase to the surface of E. coli cells. |
Databáze: | OpenAIRE |
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