Functional Dissection of the CCBE1 Protein
| Autor: | Stefan Schulte-Merker, Michael Jeltsch, Veli-Matti Leppänen, Yvonne Padberg, Kari Alitalo, Josi Peterson-Maduro, M. Guy Roukens, Frank L. Bos, Dörte Schulte |
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| Rok vydání: | 2015 |
| Předmět: |
Physiology
Vascular Endothelial Growth Factor C Mutant Regulator Craniofacial Abnormalities Mice 0302 clinical medicine vascular Gene Knock-In Techniques Lymphedema Lymphangiogenesis Non-U.S. Gov't Zebrafish Medicine(all) 0303 health sciences biology Research Support Non-U.S. Gov't Gene Expression Regulation Developmental Cell biology Phenotype Vascular endothelial growth factor C CCBE1 protein 030220 oncology & carcinogenesis Collagen Genital Diseases Male Cardiology and Cardiovascular Medicine Protein Binding Signal Transduction Endothelium vascular Genotype Protein domain Gestational Age Mice Transgenic Research Support Transfection 03 medical and health sciences Hennekam lymphangiectasia-lymphedema syndrome In vivo Journal Article Animals Humans Protein Interaction Domains and Motifs Endothelium Lymphatic Vessels 030304 developmental biology Phenocopy Binding Sites Epidermal Growth Factor Tumor Suppressor Proteins Calcium-Binding Proteins Endothelial Cells Zebrafish Proteins biology.organism_classification HEK293 Cells Mutation Immunology biology.protein Vascular endothelial growth factor Protein A Lymphangiectasis Intestinal |
| Zdroj: | Circulation Research, 116(10), 1660. Lippincott Williams and Wilkins |
| ISSN: | 1524-4571 0009-7330 |
| DOI: | 10.1161/circresaha.116.304949 |
| Popis: | Rationale : Collagen- and calcium-binding EGF domain–containing protein 1 (CCBE1) is essential for lymphangiogenesis in vertebrates and has been associated with Hennekam syndrome. Recently, CCBE1 has emerged as a crucial regulator of vascular endothelial growth factor-C (VEGFC) signaling. Objective : CCBE1 is a secreted protein characterized by 2 EGF domains and 2 collagen repeats. The functional role of the different CCBE1 protein domains is completely unknown. Here, we analyzed the functional role of the different CCBE1 domains in vivo and in vitro. Methods and Results : We analyzed the functionality of several CCBE1 deletion mutants by generating knock-in mice expressing these mutants, by analyzing their ability to enhance Vegfc signaling in vivo in zebrafish, and by testing their ability to induce VEGFC processing in vitro. We found that deleting the collagen domains of CCBE1 has a much stronger effect on CCBE1 activity than deleting the EGF domains. First, although CCBE1ΔCollagen mice fully phenocopy CCBE1 knock-out mice, CCBE1ΔEGF knock-in embryos still form rudimentary lymphatics. Second, Ccbe1ΔEGF, but not Ccbe1ΔCollagen, could partially substitute for Ccbe1 to enhance Vegfc signaling in zebrafish. Third, CCBE1ΔEGF, similarly to CCBE1, but not CCBE1ΔCollagen could activate VEGFC processing in vitro. Furthermore, a Hennekam syndrome mutation within the collagen domain has a stronger effect than a Hennekam syndrome mutation within the EGF domain. Conclusions: We propose that the collagen domains of CCBE1 are crucial for the activation of VEGFC in vitro and in vivo. The EGF domains of CCBE1 are dispensable for regulation of VEGFC processing in vitro, however, they are necessary for full lymphangiogenic activity of CCBE1 in vivo. |
| Databáze: | OpenAIRE |
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