Mouse WIF1 Is Only Modified with <i>O</i>-Fucose in Its EGF-like Domain III Despite Two Evolutionarily Conserved Consensus Sites
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 <i>O</i>-fucosylation site in its EGF-LDs III and V, the <i>O</i>...
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MDPI AG
2020-08-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/10/9/1250 |
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| author | Florian Pennarubia Emilie Pinault Bilal Al Jaam Caroline E. Brun Abderrahman Maftah Agnès Germot Sébastien Legardinier |
| author_facet | Florian Pennarubia Emilie Pinault Bilal Al Jaam Caroline E. Brun Abderrahman Maftah Agnès Germot Sébastien Legardinier |
| author_sort | Florian Pennarubia |
| collection | DOAJ |
| description | 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 <i>O</i>-fucosylation site in its EGF-LDs III and V, the <i>O</i>-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 <i>O</i>-fucosylation sites occupancy of recombinant mouse WIF1. In the monophyletic group of gnathostomes, we showed that the consensus sequence for <i>O</i>-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 <i>O</i>-fucose on its EGF-LD III. In addition, a decreased amount of mouse WIF1 in the secretome of CHO cells was observed when the <i>O</i>-fucosylation site in EGF-LD III was mutated. Based on sequence comparison and automated protein modeling, we suggest that the absence of <i>O</i>-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. |
| first_indexed | 2024-03-10T16:44:32Z |
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| language | English |
| last_indexed | 2024-03-10T16:44:32Z |
| publishDate | 2020-08-01 |
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| spelling | doaj.art-0f0ab4fe56634e00b99bb6368f67fdea2023-11-20T11:43:10ZengMDPI AGBiomolecules2218-273X2020-08-01109125010.3390/biom10091250Mouse WIF1 Is Only Modified with <i>O</i>-Fucose in Its EGF-like Domain III Despite Two Evolutionarily Conserved Consensus SitesFlorian Pennarubia0Emilie Pinault1Bilal Al Jaam2Caroline E. Brun3Abderrahman Maftah4Agnès Germot5Sébastien Legardinier6Glycosylation and cell differentiation, PEIRENE, EA 7500, Faculty of Sciences and Technology, University of Limoges, F-87060 Limoges, FranceGlycosylation and cell differentiation, PEIRENE, EA 7500, Faculty of Sciences and Technology, University of Limoges, F-87060 Limoges, FranceGlycosylation and cell differentiation, PEIRENE, EA 7500, Faculty of Sciences and Technology, University of Limoges, F-87060 Limoges, FranceGlycosylation and cell differentiation, PEIRENE, EA 7500, Faculty of Sciences and Technology, University of Limoges, F-87060 Limoges, FranceGlycosylation and cell differentiation, PEIRENE, EA 7500, Faculty of Sciences and Technology, University of Limoges, F-87060 Limoges, FranceGlycosylation and cell differentiation, PEIRENE, EA 7500, Faculty of Sciences and Technology, University of Limoges, F-87060 Limoges, FranceGlycosylation and cell differentiation, PEIRENE, EA 7500, Faculty of Sciences and Technology, University of Limoges, F-87060 Limoges, FranceThe 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 <i>O</i>-fucosylation site in its EGF-LDs III and V, the <i>O</i>-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 <i>O</i>-fucosylation sites occupancy of recombinant mouse WIF1. In the monophyletic group of gnathostomes, we showed that the consensus sequence for <i>O</i>-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 <i>O</i>-fucose on its EGF-LD III. In addition, a decreased amount of mouse WIF1 in the secretome of CHO cells was observed when the <i>O</i>-fucosylation site in EGF-LD III was mutated. Based on sequence comparison and automated protein modeling, we suggest that the absence of <i>O</i>-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.https://www.mdpi.com/2218-273X/10/9/1250click chemistryEGF-LD<i>O</i>-fucosylationphylogenyPofut1Wif1 |
| spellingShingle | Florian Pennarubia Emilie Pinault Bilal Al Jaam Caroline E. Brun Abderrahman Maftah Agnès Germot Sébastien Legardinier Mouse WIF1 Is Only Modified with <i>O</i>-Fucose in Its EGF-like Domain III Despite Two Evolutionarily Conserved Consensus Sites Biomolecules click chemistry EGF-LD <i>O</i>-fucosylation phylogeny Pofut1 Wif1 |
| title | Mouse WIF1 Is Only Modified with <i>O</i>-Fucose in Its EGF-like Domain III Despite Two Evolutionarily Conserved Consensus Sites |
| title_full | Mouse WIF1 Is Only Modified with <i>O</i>-Fucose in Its EGF-like Domain III Despite Two Evolutionarily Conserved Consensus Sites |
| title_fullStr | Mouse WIF1 Is Only Modified with <i>O</i>-Fucose in Its EGF-like Domain III Despite Two Evolutionarily Conserved Consensus Sites |
| title_full_unstemmed | Mouse WIF1 Is Only Modified with <i>O</i>-Fucose in Its EGF-like Domain III Despite Two Evolutionarily Conserved Consensus Sites |
| title_short | Mouse WIF1 Is Only Modified with <i>O</i>-Fucose in Its EGF-like Domain III Despite Two Evolutionarily Conserved Consensus Sites |
| title_sort | mouse wif1 is only modified with i o i fucose in its egf like domain iii despite two evolutionarily conserved consensus sites |
| topic | click chemistry EGF-LD <i>O</i>-fucosylation phylogeny Pofut1 Wif1 |
| url | https://www.mdpi.com/2218-273X/10/9/1250 |
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