Polylactosamine on glycoproteins influences basal levels of lymphocyte and macrophage activation

Togayachi, Akira; Kozono, Yuko; Ishida, Hiroyasu; Abe, Sumie; Suzuki, Nami; Tsunoda, Yuki; Hagiwara, Kozue; Kuno, Atsushi; Ohkura, Takashi; Sato, Nobuo; Sato, Takashi; Hirabayashi, Jun; Ikehara, Yuzuru; Tachibana, Kunihide; Narimatsu, Hisashi

doi:10.1073/pnas.0707426104

Cited by 105 publications

(103 citation statements)

References 45 publications

(37 reference statements)

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“…Immunoprecipitated materials were blotted with 5D4 or the 6B4 mouse monoclonal anti-phosphacan antibody. Lectin blot analysis for the materials was performed with LEL, which binds to polylactosamines (Togayachi et al 2007). The asterisk indicates the position of the immunoglobulin heavy chain.…”

Section: Discussionmentioning

confidence: 99%

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KSGal6ST Is Essential for the 6-Sulfation of Galactose within Keratan Sulfate in Early Postnatal Brain

Hoshino

Foyez

Ohtake-Niimi

et al. 2013

J Histochem Cytochem.

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SummaryKeratan sulfate (KS) comprises repeating disaccharides of galactose (Gal) and N-acetylglucosamine (GlcNAc). Residues of Gal and GlcNAc in KS are potentially modified with sulfate at their C-6 positions. The 5D4 monoclonal antibody recognizes KS structures containing Gal and GlcNAc, both 6-sulfated, and has been used most extensively to evaluate KS expression in mammalian brains. We previously showed that GlcNAc6ST1 is an enzyme responsible for the synthesis of the 5D4 epitope in developing brain and in the adult brain, where it is induced after injury. It has been unclear which sulfotransferase is responsible for Gal-6-sulfation within the 5D4 KS epitope in developing brains. We produced mice deficient in KSGal6ST, a Gal-6-sulfotransferase. Western blotting and immunoprecipitation revealed that all 5D4-immunoreactivity to proteins, including phosphacan, were abolished in KSGal6ST-deficient postnatal brains. Likewise, the 5D4 epitope, expressed primarily in the cortical marginal zone and subplate and dorsal thalamus, was eliminated in KSGal6ST-deficient mice. Disaccharide analysis showed the loss of Gal-6-sulfate in KS of the KSGal6ST-deficient brains. Transfection studies revealed that GlcNAc6ST1 and KSGal6ST cooperated in the expression of the 5D4 KS epitope in HeLa cells. These results indicate that KSGal6ST is essential for C-6 sulfation of Gal within KS in early postnatal brains. (J Histochem Cytochem 62:145-156, 2014)

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Section: Discussionmentioning

confidence: 99%

“…5). The 6B4 mouse monoclonal antibody and LEL, a lectin that binds to polylactosamine (Togayachi et al 2007), recognized bands in 6B4 immunoprecipitate samples of all three genotypes (Fig. 5).…”

Section: Loss Of the 5d4 Epitope In Phosphacan Expressed In P1 Brainsmentioning

confidence: 99%

KSGal6ST Is Essential for the 6-Sulfation of Galactose within Keratan Sulfate in Early Postnatal Brain

Hoshino

Foyez

Ohtake-Niimi

et al. 2013

J Histochem Cytochem.

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“…To investigate whether hematocytes manifest aberrant cell distribution or development in B3gnt5 −/− mice, we analyzed the expression of a series of CD antigens on the cell surface of peripheral blood and splenic cells using FCM. The antigens investigated were the same as those described in a previous study (19). No significant disturbance in the ratios of cell populations, such as T cells, B cells, monocytes, or granulocytes, was observed.…”

Section: Resultsmentioning

confidence: 99%

“…B3gnt2 (β3GnT2) mRNA is ubiquitously expressed in mouse tissues, and thus also in B cells, T cells, and other immune cells. The previous study demonstrated that B3gnt2 −/− B and T cells, which lack polylactosamine (PLN) on the N-glycans of their glycoproteins, showed hyperactivation following BCR or TCR/CD28 stimulation (19). Thus, PLN chains on glycoproteins have an important role in determining thresholds for in vitro immunocyte activation.…”

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confidence: 99%

Lack of lacto/neolacto-glycolipids enhances the formation of glycolipid-enriched microdomains, facilitating B cell activation

Togayachi

Kozono

Ikehara

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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In a previous study, we demonstrated that β1,3-N-acetylglucosaminyltransferase 5 (B3gnt5) is a lactotriaosylceramide (Lc 3 Cer) synthase that synthesizes a precursor structure for lacto/neolacto-series glycosphingolipids (GSLs) in in vitro experiments. Here, we generated B3gnt5-deficient (B3gnt5 −/− ) mice to investigate the in vivo biological functions of lacto/neolacto-series GSLs. In biochemical analyses, lacto/neolacto-series GSLs were confirmed to be absent and no Lc 3 Cer synthase activity was detected in the tissues of these mice. These results demonstrate that β3GnT5 is the sole enzyme synthesizing Lc 3 Cer in vivo. Ganglioside GM1, known as a glycosphingolipid-enriched microdomain (GEM) marker, was found to be upregulated in B3gnt5 −/− B cells by flow cytometry and fluorescence microscopy. However, no difference in the amount of GM1 was observed by TLC-immunoblotting analysis. The GEM-stained puncta on the surface of B3gnt5 −/− resting B cells were brighter and larger than those of WT cells. These results suggest that structural alteration of GEM occurs in B3gnt5 −/− B cells. We next examined whether BCR signaling-related proteins, such as BCR, CD19, and the signaling molecule Lyn, had moved into or out of the GEM fraction. In B3gnt5 −/− B cells, these molecules were enriched in the GEM fraction or adjacent fraction. Moreover, B3gnt5 −/− B cells were more sensitive to the induction of intracellular phosphorylation signals on BCR stimulation and proliferated more vigorously than WT B cells. Together, these results suggest that lacto/neolacto-series GSLs play an important role in clustering of GEMs and tether-specific proteins, such as BCR, CD19, and related signaling molecules to the GEMs.A lmost all organisms possess lipids and proteins to which a broad range of carbohydrate chains are linked. Some carbohydrate structures are known to participate in vital processes, such as the molecules responsible for cell-cell, receptor-ligand, and carbohydrate-carbohydrate interactions. It is known that glycosphingolipids (GSLs) have an important role in biological functions. In mammals, GSLs can be classified into several major classes, such as globo-, isoglobo-, ganglio-, and lacto/neolacto-

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“…The glycosylation of polylactosamine uses repeating Galβ1-4-glucosyl (Glc)-N-acetyl-(Ac)-β1-3 disaccharide units that are preferentially joined to β1-6GlcNAc-linked antennae that are connected to the trimannosyl core of the complex-type N-linked oligosaccharides (8). It has been reported that highly metastatic colon cancer cells synthesize more N-glycans that contain polylactosamine than cells with low metastasis (9).…”

Section: (Acetylamino)-2-deoxy-α-d-galactopyranosyl]-l-sementioning

confidence: 99%

Colon cancer cells treated with 5-fluorouracil exhibit changes in polylactosamine-type N-glycans

Gao

Shen

et al. 2014

Molecular Medicine Reports

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Abstract. 5-Fluorouracil (5-FU) is the major chemotherapeutic agent for the treatment of colorectal carcinoma, which were found to have N-glycans containing polylactosamine on the cancer cell surface. Alterations in the expression and structure of polylactosamine glycans are associated with cellular differentiation and oncogenesis. However, little is known with regard to the correlation between the levels of polylactosamine expressed in colon cancer cells and the anticancer effect of 5-FU. In the present study, SW620 cells were treated with the half maximal inhibitory concentration (IC 50 ; determined by MTT-assay) of 5-FU. Hoechst 33258 staining and flow cytometric analysis indicated that 5-FU administration resulted in apoptosis in SW620 cells. An increased percentage of cells in S phase was also observed among the SW620 cells treated with 5-FU. Under the same experimental conditions, a decrease in the 5-FU-induced inhibition of polylactosamine glycans was recorded. However, an increase in the activity of alkaline phosphatase was also observed. Furthermore, pretreatment of the SW620 cells with 5-FU inhibited the expression of β1,3-N-acetylglucosaminyltran sferase-8 (β3Gn-T8) and cluster of differentiation (CD)147 in a time-dependent manner. Overall, changes in glycosylation were associated with the anticancer effect of 5-FU in the colon cancer cells. In conclusion, polylactosamine may be a useful target for the identification of substances with anticancer activity.

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Polylactosamine on glycoproteins influences basal levels of lymphocyte and macrophage activation

Cited by 105 publications

References 45 publications

KSGal6ST Is Essential for the 6-Sulfation of Galactose within Keratan Sulfate in Early Postnatal Brain

KSGal6ST Is Essential for the 6-Sulfation of Galactose within Keratan Sulfate in Early Postnatal Brain

Lack of lacto/neolacto-glycolipids enhances the formation of glycolipid-enriched microdomains, facilitating B cell activation

Colon cancer cells treated with 5-fluorouracil exhibit changes in polylactosamine-type N-glycans

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