TY  - JOUR
A1  - Saarenpää, Tuulia
A1  - Kogan, Konstantin
A1  - Sidorova, Yulia
A1  - Mahato, Arun Kumar
A1  - Tascón, Igor
A1  - Kaljunen, Heidi
A1  - Yu, Liying
A1  - Kallijärvi, Jukka
A1  - Jurvansuu, Jaana
A1  - Saarma, Mart
A1  - Goldman, Adrian
T1  - Zebrafish GDNF and its co-receptor GFRα1 activate the human RET receptor and promote the survival of dopaminergic neurons in vitro
T2  - PLoS one
N2  - 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.
Y1  - 2017
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/43108
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-431083
SN  - 1932-6203
N1  - 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.
VL  - 12
IS  - (5): e0176166
SP  - 1
EP  - 18
PB  - PLoS
CY  - Lawrence, Kan.
ER  -