Tuulia Saarenpää, Konstantin Kogan, Yulia Sidorova, Arun Kumar Mahato, Igor Tascón, Heidi Kaljunen, Liying Yu, Jukka Kallijärvi, Jaana Jurvansuu, Mart Saarma, Adrian Goldman
- Glial cell line-derived neurotrophic factor (GDNF) is a ligand that activates, through co-receptor GDNF family receptor alpha-1 (GFRα1) and receptor tyrosine kinase “RET”, several signaling pathways crucial in the development and sustainment of multiple neuronal populations. We decided to study whether non-mammalian orthologs of these three proteins have conserved their function: can they activate the human counterparts? Using the baculovirus expression system, we expressed and purified Danio rerio RET, and its binding partners GFRα1 and GDNF, and Drosophila melanogaster RET and two isoforms of co-receptor GDNF receptor-like. Our results report high-level insect cell expression of post-translationally modified and dimerized zebrafish RET and its binding partners. We also found that zebrafish GFRα1 and GDNF are comparably active as mammalian cell-produced ones. We also report the first measurements of the affinity of the complex to RET in solution: at least for zebrafish, the Kd for GFRα1-GDNF binding RET is 5.9 μM. Surprisingly, we also found that zebrafish GDNF as well as zebrafish GFRα1 robustly activated human RET signaling and promoted the survival of cultured mouse dopaminergic neurons with comparable efficiency to mammalian GDNF, unlike E. coli-produced human proteins. These results contradict previous studies suggesting that mammalian GFRα1 and GDNF cannot bind and activate non-mammalian RET and vice versa.
MetadatenAuthor: | Tuulia Saarenpää, Konstantin Kogan, Yulia Sidorova, Arun Kumar Mahato, Igor TascónORCiD, Heidi Kaljunen, Liying Yu, Jukka Kallijärvi, Jaana Jurvansuu, Mart Saarma, Adrian Goldman |
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URN: | urn:nbn:de:hebis:30:3-431083 |
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DOI: | https://doi.org/10.1371/journal.pone.0176166 |
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ISSN: | 1932-6203 |
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Pubmed Id: | https://pubmed.ncbi.nlm.nih.gov/28467503 |
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Parent Title (German): | PLoS one |
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Publisher: | PLoS |
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Place of publication: | Lawrence, Kan. |
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Contributor(s): | Renping Zhou |
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Document Type: | Article |
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Language: | English |
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Date of Publication (online): | 2017/05/08 |
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Date of first Publication: | 2017/05/03 |
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Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
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Release Date: | 2017/05/08 |
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Volume: | 12 |
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Issue: | (5): e0176166 |
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Page Number: | 18 |
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First Page: | 1 |
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Last Page: | 18 |
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Note: | Copyright: © 2017 Saarenpää et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
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HeBIS-PPN: | 415176328 |
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Institutes: | Biochemie, Chemie und Pharmazie / Biochemie und Chemie |
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Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 59 Tiere (Zoologie) / 590 Tiere (Zoologie) |
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Sammlungen: | Universitätspublikationen |
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Licence (German): | Creative Commons - Namensnennung 4.0 |
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