Antibody-Based Fusion Proteins Allow Ca2+ Rewiring to Most Extracellular Ligands
| Autor: | Kevin Truong, Anam Qudrat |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
RHOA biology Chemistry Protein domain Biomedical Engineering General Medicine Biochemistry Genetics and Molecular Biology (miscellaneous) Fusion protein Cell biology Green fluorescent protein 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Antigen Cytoplasm biology.protein Extracellular mCherry 030217 neurology & neurosurgery |
| Zdroj: | ACS Synthetic Biology. 7:531-539 |
| ISSN: | 2161-5063 |
| Popis: | The Ca2+ signaling toolkit is the set of proteins used by living systems to generate and respond to Ca2+ signals. The selective expression of these proteins in particular tissues, cell types and subcellular locations allows the Ca2+ signal to regulate a diverse set of cellular processes. Through synthetic biology, the Ca2+ signaling toolkit can be expanded beyond the natural repertoire to potentially allow a non-natural ligand to control downstream cellular processes. To realize this potential, we exploited the ability of an antibody to bind its antigen exclusively in combination with the ability of the cytoplasmic domain of vascular endothelial growth factor receptor 2 (VEGFR2) to generate a Ca2+ signal upon oligomerization. Using protein fusions between antibody variants (i.e., nanobody, single-chain antibody and the monoclonal antibody) and the VEGFR2 cytoplasmic domain, Ca2+ signals were generated in response to extracellular stimulation with green fluorescent protein, mCherry, tumor necrosis factor alpha and soluble CD14. The Ca2+ signal generation by the stimulus did not require a stringent transition from monomer to oligomer state, but instead only required an increase in the oligomeric state. The Ca2+ signal generated by these classes of antibody-based fusion proteins can be rewired with a Ca2+ indicator or with an engineered Ca2+ activated RhoA to allow for antigen screening or migration to most extracellular ligands, respectively. |
| Databáze: | OpenAIRE |
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