Glycoproteomic analysis of the changes in protein N-glycosylation during neuronal differentiation in human-induced pluripotent stem cells and derived neuronal cells
Abstract N-glycosylation of glycoproteins, a major post-translational modification, plays a crucial role in various biological phenomena. In central nervous systems, N-glycosylation is thought to be associated with differentiation and regeneration; however, the state and role of N-glycosylation in n...
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doaj-1b5fad1276fc47159531affe4d74a5f92021-05-30T11:34:45ZengNature Publishing GroupScientific Reports2045-23222021-05-0111111210.1038/s41598-021-90102-zGlycoproteomic analysis of the changes in protein N-glycosylation during neuronal differentiation in human-induced pluripotent stem cells and derived neuronal cellsKazumasa Kimura0Takumi Koizumi1Takaya Urasawa2Yuki Ohta3Daisuke Takakura4Nana Kawasaki5Biopharmaceutical and Regenerative Sciences, Graduate School of Medical Life Science, Yokohama City UniversityBiopharmaceutical and Regenerative Sciences, Graduate School of Medical Life Science, Yokohama City UniversityBiopharmaceutical and Regenerative Sciences, Graduate School of Medical Life Science, Yokohama City UniversityBiopharmaceutical and Regenerative Sciences, Graduate School of Medical Life Science, Yokohama City UniversityBiopharmaceutical and Regenerative Sciences, Graduate School of Medical Life Science, Yokohama City UniversityBiopharmaceutical and Regenerative Sciences, Graduate School of Medical Life Science, Yokohama City UniversityAbstract N-glycosylation of glycoproteins, a major post-translational modification, plays a crucial role in various biological phenomena. In central nervous systems, N-glycosylation is thought to be associated with differentiation and regeneration; however, the state and role of N-glycosylation in neuronal differentiation remain unclear. Here, we conducted sequential LC/MS/MS analyses of tryptic digest, enriched glycopeptides, and deglycosylated peptides of proteins derived from human-induced pluripotent stem cells (iPSCs) and iPSC-derived neuronal cells, which were used as a model of neuronal differentiation. We demonstrate that the production profiles of many glycoproteins and their glycoforms were altered during neuronal differentiation. Particularly, the levels of glycoproteins modified with an N-glycan, consisting of five N-acetylhexosamines, three hexoses, and a fucose (HN5H3F), increased in dopaminergic neuron-rich cells (DAs). The N-glycan was deduced to be a fucosylated and bisected biantennary glycan based on product ion spectra. Interestingly, the HN5H3F-modified proteins were predicted to be functionally involved in neural cell adhesion, axon guidance, and the semaphorin-plexin signaling pathway, and protein modifications were site-selective and DA-selective regardless of protein production levels. Our integrated method for glycoproteome analysis and resultant profiles of glycoproteins and their glycoforms provide valuable information for further understanding the role of N-glycosylation in neuronal differentiation and neural regeneration.https://doi.org/10.1038/s41598-021-90102-z |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kazumasa Kimura Takumi Koizumi Takaya Urasawa Yuki Ohta Daisuke Takakura Nana Kawasaki |
spellingShingle |
Kazumasa Kimura Takumi Koizumi Takaya Urasawa Yuki Ohta Daisuke Takakura Nana Kawasaki Glycoproteomic analysis of the changes in protein N-glycosylation during neuronal differentiation in human-induced pluripotent stem cells and derived neuronal cells Scientific Reports |
author_facet |
Kazumasa Kimura Takumi Koizumi Takaya Urasawa Yuki Ohta Daisuke Takakura Nana Kawasaki |
author_sort |
Kazumasa Kimura |
title |
Glycoproteomic analysis of the changes in protein N-glycosylation during neuronal differentiation in human-induced pluripotent stem cells and derived neuronal cells |
title_short |
Glycoproteomic analysis of the changes in protein N-glycosylation during neuronal differentiation in human-induced pluripotent stem cells and derived neuronal cells |
title_full |
Glycoproteomic analysis of the changes in protein N-glycosylation during neuronal differentiation in human-induced pluripotent stem cells and derived neuronal cells |
title_fullStr |
Glycoproteomic analysis of the changes in protein N-glycosylation during neuronal differentiation in human-induced pluripotent stem cells and derived neuronal cells |
title_full_unstemmed |
Glycoproteomic analysis of the changes in protein N-glycosylation during neuronal differentiation in human-induced pluripotent stem cells and derived neuronal cells |
title_sort |
glycoproteomic analysis of the changes in protein n-glycosylation during neuronal differentiation in human-induced pluripotent stem cells and derived neuronal cells |
publisher |
Nature Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2021-05-01 |
description |
Abstract N-glycosylation of glycoproteins, a major post-translational modification, plays a crucial role in various biological phenomena. In central nervous systems, N-glycosylation is thought to be associated with differentiation and regeneration; however, the state and role of N-glycosylation in neuronal differentiation remain unclear. Here, we conducted sequential LC/MS/MS analyses of tryptic digest, enriched glycopeptides, and deglycosylated peptides of proteins derived from human-induced pluripotent stem cells (iPSCs) and iPSC-derived neuronal cells, which were used as a model of neuronal differentiation. We demonstrate that the production profiles of many glycoproteins and their glycoforms were altered during neuronal differentiation. Particularly, the levels of glycoproteins modified with an N-glycan, consisting of five N-acetylhexosamines, three hexoses, and a fucose (HN5H3F), increased in dopaminergic neuron-rich cells (DAs). The N-glycan was deduced to be a fucosylated and bisected biantennary glycan based on product ion spectra. Interestingly, the HN5H3F-modified proteins were predicted to be functionally involved in neural cell adhesion, axon guidance, and the semaphorin-plexin signaling pathway, and protein modifications were site-selective and DA-selective regardless of protein production levels. Our integrated method for glycoproteome analysis and resultant profiles of glycoproteins and their glycoforms provide valuable information for further understanding the role of N-glycosylation in neuronal differentiation and neural regeneration. |
url |
https://doi.org/10.1038/s41598-021-90102-z |
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