Rapid Activation of Bone Morphogenic Protein 9 by Receptor-mediated Displacement of Pro-domains*
| Autor: | Stefan Scheiblich, Mariana de Wouters, Konrad Honold, Dirk Bernicke, Yvonne Kienast, Stefan Lorenz, Christian Lehmann, Verena Brand, Martina Thier, Alexander Haas, Ute Jucknischke |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Models Molecular medicine.medical_treatment Activin Receptors Type II Recombinant Fusion Proteins GDF2 Receptors Cell Surface Biology Bone morphogenetic protein Bone Morphogenetic Protein Receptors Type II Biochemistry 03 medical and health sciences 0302 clinical medicine Antigens CD medicine Growth Differentiation Factor 2 Human Umbilical Vein Endothelial Cells Animals Humans Protein Interaction Domains and Motifs Protein Precursors Receptor Molecular Biology Cells Cultured Mice Inbred BALB C Growth factor Endoglin Growth differentiation factor Cell Biology Peptide Fragments Recombinant Proteins Cell biology Specific Pathogen-Free Organisms Growth Differentiation Factors 030104 developmental biology HEK293 Cells 030220 oncology & carcinogenesis Female Signal transduction Dimerization Transforming growth factor Signal Transduction |
| Popis: | By non-covalent association after proteolytic cleavage, the pro-domains modulate the activities of the mature growth factor domains across the transforming growth factor-β family. In the case of bone morphogenic protein 9 (BMP9), however, the pro-domains do not inhibit the bioactivity of the growth factor, and the BMP9·pro-domain complexes have equivalent biological activities as the BMP9 mature ligand dimers. By using real-time surface plasmon resonance, we could demonstrate that either binding of pro-domain-complexed BMP9 to type I receptor activin receptor-like kinase 1 (ALK1), type II receptors, co-receptor endoglin, or to mature BMP9 domain targeting antibodies leads to immediate and complete displacement of the pro-domains from the complex. Vice versa, pro-domain binding by an anti-pro-domain antibody results in release of the mature BMP9 growth factor. Based on these findings, we adjusted ELISA assays to measure the protein levels of different BMP9 variants. Although mature BMP9 and inactive precursor BMP9 protein were directly detectable by ELISA, BMP9·pro-domain complex could only be measured indirectly as dissociated fragments due to displacement of mature growth factor and pro-domains after antibody binding. Our studies provide a model in which BMP9 can be readily activated upon getting into contact with its receptors. This increases the understanding of the underlying biology of BMP9 activation and also provides guidance for ELISA development for the detection of circulating BMP9 variants. |
| Databáze: | OpenAIRE |
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