Functional Organization of Golgi N- and O-Glycosylation Pathways Involves pH-dependent Complex Formation That Is Impaired in Cancer Cells

Hassinen, Antti; Pujol, François M.; Kokkonen, Nina; Pieters, Caroline; Kihlström, Minna K.; Korhonen, Kati; Kellokumpu, Sakari

doi:10.1074/jbc.m111.277681

Cited by 112 publications

(141 citation statements)

References 40 publications

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“…It was reported that GalT-I (β-1,4-galactosyltransferase I) and ST6Gal-I (α-2,6-sialyltransferase I) form a complex that increases their enzymatic activity, suggesting that these two N-glycosyltransferases act cooperatively in N-glycan synthesis. 31 Additionally, we noticed that both galactosylation and sialylation of IgG1 tend to negatively correlate with the level of core fucosylation. IgG galactosylation and fucosylation changes going in different directions have been previously described for juvenile chronic arthritis and rheumatoid arthritis.…”

Section: ■ Discussionmentioning

confidence: 93%

High-Throughput IgG Fc N-Glycosylation Profiling by Mass Spectrometry of Glycopeptides

et al. 2013

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Age and sex dependence of subclass specific immunoglobulin G (IgG) Fc N-glycosylation was evaluated for 1709 individuals from two isolated human populations. IgGs were obtained from plasma by affinity purification using 96-well protein G monolithic plates and digested with trypsin. Fc Nglycopeptides were purified and analyzed by negative-mode MALDI-TOF-MS with 4-chloro-α-cyanocinnamic acid (Cl-CCA) matrix. Age-associated glycosylation changes were more pronounced in younger individuals (<57 years) than in older individuals (>57 years) and in females than in males. Galactosylation and sialylation decreased with increasing age and showed significant sex dependence. Interestingly, the most prominent drop in the levels of galactosylated and sialylated glycoforms in females was observed around the age of 45 to 60 years when females usually enter menopause. The incidence of bisecting N-acetylglucosamine increased in younger individuals and reached a plateau at older age. Furthermore, we compared the results to the total IgG N-glycosylation of the same populations recently analyzed by hydrophilic interaction liquid chromatography (HILIC). Significant differences were observed in the levels of galactosylation, bisecting N-acetylglucosamine and particularly sialylation, which were shown to be higher in HILIC analysis. Age and sex association of glycosylation features was, to a large extent, comparable between MALDI-TOF-MS and HILIC IgG glycosylation profiling.

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Section: ■ Discussionmentioning

confidence: 93%

High-Throughput IgG Fc N-Glycosylation Profiling by Mass Spectrometry of Glycopeptides

et al. 2013

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“…monomeric Venus (mVen), HA (YPYDVPDYA), or Myc (EQ-KLISEEDL) tag (34). Thus, the constructs for flow cytometric FRET had the tags in the C terminus of the HASs.…”

Section: Methodsmentioning

confidence: 99%

“…Immunofluorescence Microscopy and FRET Flow Cytometry-COS7, COS1, HeLa, and MCF7 cells were cultivated in DMEM as described elsewhere (34,35). One day after plating, the cells were transfected using 0.5 g of each plasmid cDNA and the FuGENE 6 TM transfection reagent (Roche Applied Science).…”

Section: Plasmids For the Microscopic Fret Analyses And Co-immunoprecmentioning

confidence: 99%

“…In each measurement, 2-5 ϫ 10 3 cells expressing both mCer and mVen were selected by gating (for setting and validation of the gates, see Ref. 34) and quantified in triplicate from three different experiments (as mCer/mVen ratios). The FRET filter set (405 ex and 515-540 em) and FloMax software were used for quantification.…”

Section: Plasmids For the Microscopic Fret Analyses And Co-immunoprecmentioning

confidence: 99%

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Fluorescence Resonance Energy Transfer (FRET) and Proximity Ligation Assays Reveal Functionally Relevant Homo- and Heteromeric Complexes among Hyaluronan Synthases HAS1, HAS2, and HAS3

Bart

Vico

Hassinen

et al. 2015

Journal of Biological Chemistry

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“…It is also possible that the kinetics of STn biosynthesis by ST6GalNAc-I is slower and takes more time to show an effect in STn expression. Alternative possibilities for aberrant STn expression in cancer contexts include mutations or promoter methylation of the molecular chaperone Cosmc, 38,39 aberrant localization of glycosyltransferases in the ER/Golgi, 40,41 or the activity of another sialyltransferase from the same sub-family as ST6GalNAc-I-ST6GalNAc-II 4 -have been demonstrated. This enzyme, ST6GalNAc-II, was also shown to have some specificity in vitro for GalNAc substrates O-linked to proteins (Tn antigen).…”

Section: Cdx2 Regulates St6galnac-i In Intestinal Metaplasia R Pinto mentioning

confidence: 99%

CDX2 homeoprotein is involved in the regulation of ST6GalNAc-I gene in intestinal metaplasia

Pinto

Barros

Pereira-Castro

et al. 2015

Laboratory Investigation

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De novo expression of Sialyl-Tn (STn) antigen is one of the most common features of intestinal metaplasia (IM) and gastric carcinomas, and its biosynthesis has been mostly attributed to ST6GalNAc-I activity. However, the regulation of this glycosyltransferase expression is not elucidated. In IM lesions and in the intestine, CDX2 homeobox transcription factor is co-expressed with STn and ST6GalNAc-I. We therefore hypothesized that CDX2 might induce STn expression by positive regulation of ST6GalNAc-I. We showed that ST6GalNAc-I transcript levels and CDX2 have a coordinated expression upon Caco-2 in vitro differentiation, and overexpression of CDX2 in MKN45 gastric cells increases ST6GalNAc-I transcript levels. Nine putative CDX-binding sites in the ST6GalNAc-I-regulatory sequence were identified and analyzed by chromatin immunoprecipitation in Caco-2 cells and in IM. The results showed that CDX2 protein is recruited to all regions, being the most proximal sites preferentially occupied in vivo. Luciferase assays demonstrated that CDX2 is able to transactivate ST6GalNac-I-regulatory region. The induction was stronger for the regions mapped in the neighbourhood of ATG start codon and site-directed mutagenesis of these sites confirmed their importance. In conclusion, we show that CDX2 transcriptionally regulates ST6GalNAc-I gene expression, specifically in the preneoplastic IM lesion.

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Functional Organization of Golgi N- and O-Glycosylation Pathways Involves pH-dependent Complex Formation That Is Impaired in Cancer Cells

Cited by 112 publications

References 40 publications

High-Throughput IgG Fc N-Glycosylation Profiling by Mass Spectrometry of Glycopeptides

High-Throughput IgG Fc N-Glycosylation Profiling by Mass Spectrometry of Glycopeptides

Fluorescence Resonance Energy Transfer (FRET) and Proximity Ligation Assays Reveal Functionally Relevant Homo- and Heteromeric Complexes among Hyaluronan Synthases HAS1, HAS2, and HAS3

CDX2 homeoprotein is involved in the regulation of ST6GalNAc-I gene in intestinal metaplasia

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