Fibulin-5 Regulates Angiopoietin-1/Tie-2 Receptor Signaling in Endothelial Cells
| Autor: | Sabah N. A. Hussain, Kerstin Tiedemann, Veronica Sanchez, Hodan Ismail, Dominique Mayaki, Wilson Ventura Chan, Elaine C. Davis |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Inhibitor of Differentiation Protein 1 Integrins lcsh:Medicine Gene Expression Plasma protein binding Biochemistry Epithelium Binding Analysis 0302 clinical medicine Animal Cells Cricetinae Medicine and Health Sciences Recombinant Protein Purification Enzyme-Linked Immunoassays Post-Translational Modification Phosphorylation lcsh:Science Cells Cultured Extracellular Matrix Proteins Multidisciplinary Reverse Transcriptase Polymerase Chain Reaction Receptor TIE-2 Recombinant Proteins Cell biology Angiogenesis inhibitor Extracellular Matrix Endothelial stem cell 030220 oncology & carcinogenesis KLF2 Signal transduction Cellular Types Anatomy Cellular Structures and Organelles Research Article Protein Binding Signal Transduction Protein Purification Cell Survival Immunoblotting Kruppel-Like Transcription Factors Molecular Probe Techniques CHO Cells Biology Research and Analysis Methods 03 medical and health sciences Vasculogenesis Cricetulus Cell Adhesion Angiopoietin-1 Human Umbilical Vein Endothelial Cells Animals Humans Immunoassays Molecular Biology Techniques Transcription factor Molecular Biology Chemical Characterization Early Growth Response Protein 1 HEK 293 cells lcsh:R Biology and Life Sciences Endothelial Cells Proteins Epithelial Cells Cell Biology Molecular biology 030104 developmental biology Biological Tissue HEK293 Cells Mutation Immunologic Techniques lcsh:Q Purification Techniques |
| Zdroj: | PLoS ONE PLoS ONE, Vol 11, Iss 6, p e0156994 (2016) |
| ISSN: | 1932-6203 |
| Popis: | Background Fibulin-5 is an extracellular matrix glycoprotein that plays critical roles in vasculogenesis and embryonic development. Deletion of Fibulin-5 in mice results in enhanced skin vascularization and upregulation of the angiogenesis factor angiopoietin-1 (Ang-1), suggesting that Fibulin-5 functions as an angiogenesis inhibitor. In this study, we investigate the inhibitory effects of Fibulin-5 on Ang-1/TIE-2 receptor pathway signaling and cell survival in human endothelial cells. Methodology/Principal Findings Recombinant wild-type and RGE-mutant Fibulin-5 proteins were generated through stable transfection of HEK293 and CHO cells, respectively. In vitro solid phase binding assays using pure proteins revealed that wild-type Fibulin-5 does not bind to Ang-1 or TIE-2 proteins but strongly binds to heparin. Binding assays using human umbilical vein endothelial cells (HUVECs) indicated that wild-type Fibulin-5 strongly binds to cells but RGE-mutant Fibulin-5, which is incapable of binding to integrins, does not. Pre-incubation of HUVECs for 1 hr with Fibulin-5 significantly increased caspase 3/7 activity, ERK1/2 phosphorylation, and expressions of the transcription factor early growth response 1 (EGR1) and the dual-specificity phosphatase 5 (DUSP5). Fibulin-5 also strongly attenuated Ang-1-induced TIE-2 and AKT phosphorylation, decreased Ang-1-induced expressions of the transcription factors Inhibitor of DNA Binding 1 (ID1) and Kruppel-like Factor 2 (KLF2), and reversed the inhibitory effect of Ang-1 on serum deprivation-induced cytotoxicity and caspase 3/7 activity. Conclusion/Significance We conclude that Fibulin-5 strongly binds to the endothelial cell surface through heparin-sulfate proteoglycans and possibly integrins and that it exerts strong anti-angiogenic effects by reducing endothelial cell viability and interfering with the signaling pathways of the Ang-1/TIE-2 receptor axis. |
| Databáze: | OpenAIRE |
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