Metabolic Reprogramming by Hexosamine Biosynthetic and Golgi N-Glycan Branching Pathways

Ryczko, Michael; Pawling, Judy; Chen, Rui; Rahman, Anas M. Abdel; Yau, Kevin; Copeland, John W.; Zhang, Cunjie; Surendra, Anu; Guttman, David S.; Figeys, Daniel; Dennis, James W.

doi:10.1038/srep23043

Cited by 90 publications

(91 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…GlcNAc has recently been reported to promote glucose, glutamine, and fatty acid uptake in liver and/or adipose tissue via branching induced increases in surface retention of metabolite transporters (Ryczko et al, 2016). Similarly, we observed that modifying branching in T cells with GlcNAc or kifunensine altered glucose uptake.…”

Section: Discussionmentioning

confidence: 99%

Glycolysis and glutaminolysis cooperatively control T cell function by limiting metabolite supply to N-glycosylation

Araujo

Khim

Mkhikian

et al. 2017

eLife

170

183

View full text Add to dashboard Cite

Rapidly proliferating cells switch from oxidative phosphorylation to aerobic glycolysis plus glutaminolysis, markedly increasing glucose and glutamine catabolism. Although Otto Warburg first described aerobic glycolysis in cancer cells >90 years ago, the primary purpose of this metabolic switch remains controversial. The hexosamine biosynthetic pathway requires glucose and glutamine for de novo synthesis of UDP-GlcNAc, a sugar-nucleotide that inhibits receptor endocytosis and signaling by promoting N-acetylglucosamine branching of Asn (N)-linked glycans. Here, we report that aerobic glycolysis and glutaminolysis co-operatively reduce UDP-GlcNAc biosynthesis and N-glycan branching in mouse T cell blasts by starving the hexosamine pathway of glucose and glutamine. This drives growth and pro-inflammatory TH17 over anti-inflammatory-induced T regulatory (iTreg) differentiation, the latter by promoting endocytic loss of IL-2 receptor-α (CD25). Thus, a primary function of aerobic glycolysis and glutaminolysis is to co-operatively limit metabolite supply to N-glycan biosynthesis, an activity with widespread implications for autoimmunity and cancer.DOI: http://dx.doi.org/10.7554/eLife.21330.001

show abstract

Section: Discussionmentioning

confidence: 99%

Glycolysis and glutaminolysis cooperatively control T cell function by limiting metabolite supply to N-glycosylation

Araujo

Khim

Mkhikian

et al. 2017

eLife

170

183

View full text Add to dashboard Cite

show abstract

“…The presence of the β(1,6)GlcNAc branch on N-glycans is readily detected by enhanced binding of the plant lectin L-PHA to glycoproteins, and cannot occur on hybrid N-glycans. It has most recently been shown that by increasing GlcNAc levels in cells and mice, compensation of N-glycan levels may occur via enhanced metabolism and increased UDP-GlcNAc levels in Mgat5 null mice [74]. In the absence of MGAT5, the glucagon receptor in liver and hepatocytes is functionally impaired, and hexosamine biosynthesis is reduced [75].…”

Section: Pathway-specific Glycans That Affect Postnatal Developmentmentioning

confidence: 99%

What Have We Learned from Glycosyltransferase Knockouts in Mice?

Stanley

2016

Journal of Molecular Biology

View full text Add to dashboard Cite

There are five major classes of glycan including N- and O-glycans, glycosaminoglycans, glycosphingolipids and glycophosphatidylinositol anchors, all expressed at the molecular frontier of each mammalian cell. Numerous biological consequences of altering the expression of mammalian glycans are understood at a mechanistic level, but many more remain to be characterized. Mouse mutants with deleted, defective, or misexpressed genes that encode activities necessary for glycosylation have led the way to identifying key functions of glycans in biology. However, with the advent of exome sequencing, humans with mutations in genes involved in glycosylation are also revealing specific requirements for glycans in mammalian development. The aim of this review is to summarize glycosylation genes that are necessary for mouse embryonic development; pathway-specific glycosylation genes whose deletion leads to postnatal morbidity; and glycosylation genes for which effects are mild, but perturbation of the organism may reveal functional consequences. General strategies for generating and interpreting the phenotype of mice with glycosylation defects are discussed in relation to human congenital disorders of glycosylation (CDG).

show abstract

“…Altering glycostructures in production processes is mostly done by variation of production conditions or by changing the activity level of specific glycosyltransferases and glycosyl hydrolases and has primarily been studied on glycoproteins with a single N-glycosylation site (monoclonal antibody) (22,(29)(30)(31)(32)(33). The analysis of site-specific microheterogeneity on a protein with multiple N-glycosylation sites in cell or tissue-derived extracts requires more elaborate analytical methods that have become available only recently (16,17,(34)(35)(36).…”

mentioning

confidence: 99%

Influence of protein/glycan interaction on site‐specific glycan heterogeneity

Losfeld¹,

Scibona²,

Lin³

et al. 2017

FASEB j.

View full text Add to dashboard Cite

To study how the interaction between N-linked glycans and the surrounding amino acids influences oligosaccharide processing, we used protein disulfide isomerase (PDI), a glycoprotein bearing 5 N-glycosylation sites, as a model system and expressed it transiently in a Chinese hamster ovary (CHO)-S cell line. PDI was produced as both secreted Sec-PDI and endoplasmic reticulum-retained glycoprotein (ER)-PDI, to study glycan processing by ER and Golgi resident enzymes. Quantitative site-specific glycosylation profiles were obtained, and flux analysis enabled modeling site-specific glycan processing. By altering the primary sequence of PDI, we changed the glycan/ protein interaction and thus the site-specific glycoprofile because of the improved enzymatic fluxes at enzymatic bottlenecks. Our results highlight the importance of direct interactions between N-glycans and surface-exposed amino acids of glycoproteins on processing in the ER and the Golgi and the possibility of changing a site-specific N-glycan profile by modulating such interactions and thus the associated enzymatic fluxes. Altering the primary protein sequence can therefore be used to glycoengineer recombinant proteins.-Losfeld, M.-E., Scibona, E., Lin, C.-W., Villiger, T. K., Gauss, R., Morbidelli, M., Aebi, M. Influence of protein/glycan interaction on site-specific glycan heterogeneity. FASEB J. 31, 4623-4635 (2017). www.fasebj.org KEY WORDS: glycoengineering • glycobiology • glycoprotein maturation • Golgi processing • enzymatic flux N-glycosylation is a major posttranslational modification of proteins that modulates folding, maturation, trafficking, and secretion, as well as specific protein functions (1-3). Contrary to many other posttranslational modifications, N-glycans are composed of many monosaccharide units, creating a large diversity of glycan structures (4, 5). In eukaryotic cells, the N-glycosylation occurs in the endoplasmic reticulum (ER) after a conserved pathway. The oligosaccharide GlcNAc 2 Man 9 Glc 3 is assembled on the lipid dolicholpyrophosphate at the membrane of the ER and then is covalently linked to the side-chain amide of the asparagine residue in the sequon N-X-S/T by oligosaccharyltransferase (OST) (1, 6, 7). Multiple sites on one glycoprotein can be modified by the OST complex and with the increasing complexity of multicellular organism, an increasing number of N-glycosylation sites in the glycoproteome are observed. It is estimated that up to 6000 different sites are modified by OST in mammalian cells (8). In the second phase of the N-glycosylation pathway, the protein-linked glycan is trimmed by glucosidases and mannosidases (1, 9) and is subsequently modified by different glycosyltransferases spatially distributed along the secretory pathway. N-glycan processing enzymes have defined substrate specificities and locations in the secretory pathway (9, 10), and their expression level can be regulated upon internal and external signals.Glycan maturation in the secretory pathway is not a template-driven process. The...

show abstract

Metabolic Reprogramming by Hexosamine Biosynthetic and Golgi N-Glycan Branching Pathways

Cited by 90 publications

References 63 publications

Glycolysis and glutaminolysis cooperatively control T cell function by limiting metabolite supply to N-glycosylation

Glycolysis and glutaminolysis cooperatively control T cell function by limiting metabolite supply to N-glycosylation

What Have We Learned from Glycosyltransferase Knockouts in Mice?

Influence of protein/glycan interaction on site‐specific glycan heterogeneity

Contact Info

Product

Resources

About