Zebrafish GDNF and its co-receptor GFRα1 activate the human RET receptor and promote the survival of dopaminergic neurons in vitro.
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...
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Public Library of Science (PLoS)
2017-01-01
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| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0176166&type=printable |
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| author | Tuulia Saarenpää Konstantin Kogan Yulia Sidorova Arun Kumar Mahato Igor Tascón Heidi Kaljunen Liying Yu Jukka Kallijärvi Jaana Jurvansuu Mart Saarma Adrian Goldman |
| author_facet | Tuulia Saarenpää Konstantin Kogan Yulia Sidorova Arun Kumar Mahato Igor Tascón Heidi Kaljunen Liying Yu Jukka Kallijärvi Jaana Jurvansuu Mart Saarma Adrian Goldman |
| author_sort | Tuulia Saarenpää |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-1bf8ffc84a4e44968a0e05cf175973f4 |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-1bf8ffc84a4e44968a0e05cf175973f42025-08-20T02:31:59ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01125e017616610.1371/journal.pone.0176166Zebrafish GDNF and its co-receptor GFRα1 activate the human RET receptor and promote the survival of dopaminergic neurons in vitro.Tuulia SaarenpääKonstantin KoganYulia SidorovaArun Kumar MahatoIgor TascónHeidi KaljunenLiying YuJukka KallijärviJaana JurvansuuMart SaarmaAdrian GoldmanGlial 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.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0176166&type=printable |
| spellingShingle | Tuulia Saarenpää Konstantin Kogan Yulia Sidorova Arun Kumar Mahato Igor Tascón Heidi Kaljunen Liying Yu Jukka Kallijärvi Jaana Jurvansuu Mart Saarma Adrian Goldman Zebrafish GDNF and its co-receptor GFRα1 activate the human RET receptor and promote the survival of dopaminergic neurons in vitro. PLoS ONE |
| title | Zebrafish GDNF and its co-receptor GFRα1 activate the human RET receptor and promote the survival of dopaminergic neurons in vitro. |
| title_full | Zebrafish GDNF and its co-receptor GFRα1 activate the human RET receptor and promote the survival of dopaminergic neurons in vitro. |
| title_fullStr | Zebrafish GDNF and its co-receptor GFRα1 activate the human RET receptor and promote the survival of dopaminergic neurons in vitro. |
| title_full_unstemmed | Zebrafish GDNF and its co-receptor GFRα1 activate the human RET receptor and promote the survival of dopaminergic neurons in vitro. |
| title_short | Zebrafish GDNF and its co-receptor GFRα1 activate the human RET receptor and promote the survival of dopaminergic neurons in vitro. |
| title_sort | zebrafish gdnf and its co receptor gfrα1 activate the human ret receptor and promote the survival of dopaminergic neurons in vitro |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0176166&type=printable |
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