title: Engineering Synthetic Proteins to Generate Ca2+Signals in Mammalian Cells.
abstract: The versatility of Ca2+signals allows it to regulate diverse cellular processes such as migration, apoptosis, motility and exocytosis. In some receptors (e.g., VEGFR2), Ca2+signals are generated upon binding their ligand(s) (e.g., VEGF-A). Here, we employed a design strategy to engineer proteins that generate a Ca2+signal upon binding various extracellular stimuli by creating fusions of protein domains that oligomerize to the transmembrane domain and the cytoplasmic tail of the VEGFR2. To test the strategy, we created chimeric proteins that generate Ca2+signals upon stimulation with various extracellular stimuli (e.g., rapamycin, EDTA or extracellular free Ca2+). By coupling these chimeric proteins that generate Ca2+signals with proteins that respond to Ca2+signals, we rewired, for example, dynamic cellular blebbing to increases in extracellular free Ca2+. Thus, using this design strategy, it is possible to engineer proteins to generate a Ca2+signal to rewire a wide range of extracellular stimuli to a wide range of Ca2+-activated processes.
authors.affiliation: Institute of Biomaterials and Biomedical Engineering, University of Toronto , 164 College Street, Toronto, Ontario M5S 3G9, Canada.
authors.forename: Anam
authors.initials: A
authors.lastname: Qudrat
authors.affiliation: Institute of Biomaterials and Biomedical Engineering, University of Toronto , 164 College Street, Toronto, Ontario M5S 3G9, Canada.|Edward S. Rogers, Sr. Department of Electrical and Computer Engineering, University of Toronto , 10 King's College Circle, Toronto, Ontario M5S 3G4, Canada.
authors.forename: Kevin
authors.identifier: 0000-0002-9520-2144
authors.initials: K
authors.lastname: Truong
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008322:Mammals / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D011506:Proteins; D040301:Vascular Endothelial Growth Factor Receptor-2; D002118:Calcium
keywords: Ca2+ signaling; VEGFR2; chimeras; plasma membrane; protein engineering; synthetic biology; transmembrane proteins