5 years ago

Impact of the Niemann–Pick c1 Gene Mutation on the Total Cellular Glycomics of CHO Cells

Impact of the Niemann–Pick c1 Gene Mutation on the Total Cellular Glycomics of CHO Cells
Jun-ichi Furukawa, Yasuro Shinohara, Tetsumi Irie, Kazue Okada, Jinhua Piao, Ikuko Yokota, Takumi Era, Minami Soga
Niemann–Pick disease type C (NPC) is an autosomal recessive lipid storage disorder, and the majority of cases are caused by mutations in the NPC1 gene. In this study, we clarified how a single gene mutation in the NPC1 gene impacts the cellular glycome by analyzing the total glycomic expression profile of Chinese hamster ovary cell mutants defective in the Npc1 gene (Npc1 KO CHO cells). A number of glycomic alterations were identified, including increased expression of lactosylceramide, GM1, GM2, GD1, various neolacto-series glycosphingolipids, and sialyl-T (O-glycan), which was found to be the major sialylated protein-bound glycan, as well as various N-glycans, which were commonly both fucosylated and sialylated. We also observed significant increases in the total amounts of free oligosaccharides (fOSs), especially in the unique complex- and hybrid-type fOSs. Treatment of Npc1 KO CHO cells with 2-hydroxypropyl-β-cyclodextrin (HPBCD), which can reduce cholesterol and glycosphingolipid (GSL) storage, did not affect the glycomic alterations observed in the GSL-, N-, and O-glycans of Npc1 KO CHO cells. However, HPBCD treatment corrected the glycomic alterations observed in fOSs to levels observed in wild-type cells.

Publisher URL: http://dx.doi.org/10.1021/acs.jproteome.7b00070

DOI: 10.1021/acs.jproteome.7b00070

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