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Mouse WIF1 Is Only Modified with O-Fucose in Its EGF-like Domain III Despite Two Evolutionarily Conserved Consensus Sites
- Source :
- Biomolecules, Vol 10, Iss 1250, p 1250 (2020), Biomolecules, Volume 10, Issue 9
- Publication Year :
- 2020
- Publisher :
- MDPI AG, 2020.
-
Abstract
- The Wnt Inhibitory Factor 1 (Wif1), known to inhibit Wnt signaling pathways, is composed of a WIF domain and five EGF-like domains (EGF-LDs) involved in protein interactions. Despite the presence of a potential O-fucosylation site in its EGF-LDs III and V, the O-fucose sites occupancy has never been demonstrated for WIF1. In this study, a phylogenetic analysis on the distribution, conservation and evolution of Wif1 proteins was performed, as well as biochemical approaches focusing on O-fucosylation sites occupancy of recombinant mouse WIF1. In the monophyletic group of gnathostomes, we showed that the consensus sequence for O-fucose modification by Pofut1 is highly conserved in Wif1 EGF-LD III while it was more divergent in EGF-LD V. Using click chemistry and mass spectrometry, we demonstrated that mouse WIF1 was only modified with a non-extended O-fucose on its EGF-LD III. In addition, a decreased amount of mouse WIF1 in the secretome of CHO cells was observed when the O-fucosylation site in EGF-LD III was mutated. Based on sequence comparison and automated protein modeling, we suggest that the absence of O-fucose on EGF-LD V of WIF1 in mouse and probably in most gnathostomes, could be related to EGF-LD V inability to interact with POFUT1.
- Subjects :
- EGF-like domain
lcsh:QR1-502
WIF1
phylogeny
Biochemistry
Fucose
lcsh:Microbiology
Protein–protein interaction
law.invention
03 medical and health sciences
chemistry.chemical_compound
Pofut1
law
EGF-LD
Consensus sequence
Molecular Biology
030304 developmental biology
0303 health sciences
Chinese hamster ovary cell
030302 biochemistry & molecular biology
Wnt signaling pathway
Cell biology
chemistry
click chemistry
Recombinant DNA
O-fucosylation
Wif1
hormones, hormone substitutes, and hormone antagonists
Subjects
Details
- Language :
- English
- Volume :
- 10
- Issue :
- 1250
- Database :
- OpenAIRE
- Journal :
- Biomolecules
- Accession number :
- edsair.doi.dedup.....ba399311c19c112035e7c8c09a701b27