Formation and characterization of BMP2/GDF5 and BMP4/GDF5 heterodimers
Abstract Background Proteins of the TGFβ family, which are largely studied as homodimers, are also known to form heterodimers with biological activity distinct from their component homodimers. For instance, heterodimers of bone morphogenetic proteins, including BMP2/BMP7, BMP2/BMP6, and BMP9/BMP10,...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-02-01
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| Series: | BMC Biology |
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| Online Access: | https://doi.org/10.1186/s12915-023-01522-4 |
| _version_ | 1811171570577571840 |
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| author | Gregory R. Gipson Kristof Nolan Chandramohan Kattamuri Alan P. Kenny Zachary Agricola Nicole A. Edwards Joseph Zinski Magdalena Czepnik Mary C. Mullins Aaron M. Zorn Thomas B. Thompson |
| author_facet | Gregory R. Gipson Kristof Nolan Chandramohan Kattamuri Alan P. Kenny Zachary Agricola Nicole A. Edwards Joseph Zinski Magdalena Czepnik Mary C. Mullins Aaron M. Zorn Thomas B. Thompson |
| author_sort | Gregory R. Gipson |
| collection | DOAJ |
| description | Abstract Background Proteins of the TGFβ family, which are largely studied as homodimers, are also known to form heterodimers with biological activity distinct from their component homodimers. For instance, heterodimers of bone morphogenetic proteins, including BMP2/BMP7, BMP2/BMP6, and BMP9/BMP10, among others, have illustrated the importance of these heterodimeric proteins within the context of TGFβ signaling. Results In this study, we have determined that mature GDF5 can be combined with mature BMP2 or BMP4 to form BMP2/GDF5 and BMP4/GDF5 heterodimer. Intriguingly, this combination of a BMP2 or BMP4 monomer, which exhibit high affinity to heparan sulfate characteristic to the BMP class, with a GDF5 monomer with low heparan sulfate affinity produces a heterodimer with an intermediate affinity. Using heparin affinity chromatography to purify the heterodimeric proteins, we then determined that both the BMP2/GDF5 and BMP4/GDF5 heterodimers consistently signaled potently across an array of cellular and in vivo systems, while the activities of their homodimeric counterparts were more context dependent. These differences were likely driven by an increase in the combined affinities for the type 1 receptors, Alk3 and Alk6. Furthermore, the X-ray crystal structure of BMP2/GDF5 heterodimer was determined, highlighting the formation of two asymmetric type 1 receptor binding sites that are both unique relative to the homodimers. Conclusions Ultimately, this method of heterodimer production yielded a signaling molecule with unique properties relative to the homodimeric ligands, including high affinity to multiple type 1 and moderate heparan binding affinity. |
| first_indexed | 2024-04-10T17:16:14Z |
| format | Article |
| id | doaj.art-777da4f305fb4a5cbabba5e6f768a74b |
| institution | Directory Open Access Journal |
| issn | 1741-7007 |
| language | English |
| last_indexed | 2024-04-10T17:16:14Z |
| publishDate | 2023-02-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biology |
| spelling | doaj.art-777da4f305fb4a5cbabba5e6f768a74b2023-02-05T12:23:50ZengBMCBMC Biology1741-70072023-02-0121112010.1186/s12915-023-01522-4Formation and characterization of BMP2/GDF5 and BMP4/GDF5 heterodimersGregory R. Gipson0Kristof Nolan1Chandramohan Kattamuri2Alan P. Kenny3Zachary Agricola4Nicole A. Edwards5Joseph Zinski6Magdalena Czepnik7Mary C. Mullins8Aaron M. Zorn9Thomas B. Thompson10Department of Molecular & Cellular Biosciences, University of Cincinnati College of MedicineDepartment of Biochemistry and Molecular Biophysics, University of ChicagoDepartment of Molecular & Cellular Biosciences, University of Cincinnati College of MedicinePerinatal Institute, Divisions of Developmental Biology and Neonatology & Pulmonary Biology, Cincinnati Children’s Hospital Medical CenterPerinatal Institute, Divisions of Developmental Biology and Neonatology & Pulmonary Biology, Cincinnati Children’s Hospital Medical CenterPerinatal Institute, Divisions of Developmental Biology and Neonatology & Pulmonary Biology, Cincinnati Children’s Hospital Medical CenterDepartment of Cell and Developmental Biology, University of Pennsylvania Perelman School of MedicineDepartment of Molecular & Cellular Biosciences, University of Cincinnati College of MedicineDepartment of Cell and Developmental Biology, University of Pennsylvania Perelman School of MedicinePerinatal Institute, Divisions of Developmental Biology and Neonatology & Pulmonary Biology, Cincinnati Children’s Hospital Medical CenterDepartment of Molecular & Cellular Biosciences, University of Cincinnati College of MedicineAbstract Background Proteins of the TGFβ family, which are largely studied as homodimers, are also known to form heterodimers with biological activity distinct from their component homodimers. For instance, heterodimers of bone morphogenetic proteins, including BMP2/BMP7, BMP2/BMP6, and BMP9/BMP10, among others, have illustrated the importance of these heterodimeric proteins within the context of TGFβ signaling. Results In this study, we have determined that mature GDF5 can be combined with mature BMP2 or BMP4 to form BMP2/GDF5 and BMP4/GDF5 heterodimer. Intriguingly, this combination of a BMP2 or BMP4 monomer, which exhibit high affinity to heparan sulfate characteristic to the BMP class, with a GDF5 monomer with low heparan sulfate affinity produces a heterodimer with an intermediate affinity. Using heparin affinity chromatography to purify the heterodimeric proteins, we then determined that both the BMP2/GDF5 and BMP4/GDF5 heterodimers consistently signaled potently across an array of cellular and in vivo systems, while the activities of their homodimeric counterparts were more context dependent. These differences were likely driven by an increase in the combined affinities for the type 1 receptors, Alk3 and Alk6. Furthermore, the X-ray crystal structure of BMP2/GDF5 heterodimer was determined, highlighting the formation of two asymmetric type 1 receptor binding sites that are both unique relative to the homodimers. Conclusions Ultimately, this method of heterodimer production yielded a signaling molecule with unique properties relative to the homodimeric ligands, including high affinity to multiple type 1 and moderate heparan binding affinity.https://doi.org/10.1186/s12915-023-01522-4BMPGrowth factorHeterodimerCell signaling |
| spellingShingle | Gregory R. Gipson Kristof Nolan Chandramohan Kattamuri Alan P. Kenny Zachary Agricola Nicole A. Edwards Joseph Zinski Magdalena Czepnik Mary C. Mullins Aaron M. Zorn Thomas B. Thompson Formation and characterization of BMP2/GDF5 and BMP4/GDF5 heterodimers BMC Biology BMP Growth factor Heterodimer Cell signaling |
| title | Formation and characterization of BMP2/GDF5 and BMP4/GDF5 heterodimers |
| title_full | Formation and characterization of BMP2/GDF5 and BMP4/GDF5 heterodimers |
| title_fullStr | Formation and characterization of BMP2/GDF5 and BMP4/GDF5 heterodimers |
| title_full_unstemmed | Formation and characterization of BMP2/GDF5 and BMP4/GDF5 heterodimers |
| title_short | Formation and characterization of BMP2/GDF5 and BMP4/GDF5 heterodimers |
| title_sort | formation and characterization of bmp2 gdf5 and bmp4 gdf5 heterodimers |
| topic | BMP Growth factor Heterodimer Cell signaling |
| url | https://doi.org/10.1186/s12915-023-01522-4 |
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