PUBLICATION
Zebrafish GDNF and its co-receptor GFR?1 activate the human RET receptor and promote the survival of dopaminergic neurons in vitro
- Authors
- Saarenpää, T., Kogan, K., Sidorova, Y., Mahato, A.K., Tascón, I., Kaljunen, H., Yu, L., Kallijärvi, J., Jurvansuu, J., Saarma, M., Goldman, A.
- ID
- ZDB-PUB-170504-3
- Date
- 2017
- Source
- PLoS One 12: e0176166 (Journal)
- Registered Authors
- Keywords
- Zebrafish, Neurons, Frogs, Phosphorylation, MAPK signaling cascades, Dopaminergics, Luciferase assay, Elution
- MeSH Terms
-
- Glial Cell Line-Derived Neurotrophic Factor Receptors/chemistry
- Glial Cell Line-Derived Neurotrophic Factor Receptors/metabolism*
- Phosphorylation
- Humans
- Sequence Homology, Amino Acid
- Cell Survival
- Proto-Oncogene Proteins c-ret/metabolism*
- Glial Cell Line-Derived Neurotrophic Factor/metabolism*
- Dopamine/metabolism*
- Amino Acid Sequence
- Zebrafish
- Neurons/metabolism*
- Animals
- PubMed
- 28467503 Full text @ PLoS One
Citation
Saarenpää, T., Kogan, K., Sidorova, Y., Mahato, A.K., Tascón, I., Kaljunen, H., Yu, L., Kallijärvi, J., Jurvansuu, J., Saarma, M., Goldman, A. (2017) Zebrafish GDNF and its co-receptor GFR?1 activate the human RET receptor and promote the survival of dopaminergic neurons in vitro. PLoS One. 12:e0176166.
Abstract
Glial cell line-derived neurotrophic factor (GDNF) is a ligand that activates, through co-receptor GDNF family receptor alpha-1 (GFR?1) 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 GFR?1 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 GFR?1 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 GFR?1-GDNF binding RET is 5.9 ?M. Surprisingly, we also found that zebrafish GDNF as well as zebrafish GFR?1 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 GFR?1 and GDNF cannot bind and activate non-mammalian RET and vice versa.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping