Cloning, expression and gene organization of a human Neu5Acα2‒3Galβ1‒3GalNAc α2,6-sialyltransferase: hST6GalNAc IV

Harduin-Lepers, Anne; Stokes, David C.; Steelant, Wim F.A.; Samyn-Petit, Bénédicte; Krzewinski-Recchi, Marie-Ange; Vallejo-Ruíz, Verónica; Zanetta, Jean‐Pierre; Augé, Claudine; Delannoy, Philippe

doi:10.1042/0264-6021:3520037

Cited by 28 publications

(25 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Among these sialyltransferases, the ST6GalNAc IV gene was cloned and characterized from the human and mouse, but not from pig [3,11]. However, pig pST6GalNAc IV gene has only been reported from the NCBI Genebank database (GenBank accession number EU643700 and AJ620652).…”

Section: Discussionmentioning

confidence: 98%

“…It was reported that human ST6GalNAc IV was expressed at low level in most human tissues and human cancer cell lines [11]. Furthermore, the mouse ST6GalNAc IV was specifically expressed in brain, colon and generally expressed in lung, thymus and spleen [3].…”

Section: Discussionmentioning

confidence: 98%

“…So far, six types of GalNAc α2,6-sialyltransferase including ST6GalNAc I, II, III, IV, V and VI have been reported [3,[11][12][13][14][15]. Among these GalNAc α2,6-sialyltransferase family, the cDNA of ST6GalNAc IV has been isolated from human and mouse [3,11].…”

Section: Introductionmentioning

confidence: 98%

“…Among these GalNAc α2,6-sialyltransferase family, the cDNA of ST6GalNAc IV has been isolated from human and mouse [3,11]. It was demonstrated that ST6GalNAc IV has restricted substrate specificity which utilizes Neu5Acα2-3Galβ1-3GalNAc trisaccharides found on O-glycosylated proteins [1,3].…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Molecular characterization of pig α2,3-Gal-β1,3-GalNAc-α2,6-sialyltransferase (pST6GalNAc IV) gene specific for Neu5Acα2-3Galβ1-3GalNAc trisaccharide structure

Song

Jin

et al. 2010

Glycoconj J

View full text Add to dashboard Cite

Sialic acids of glycoconjugates play crucial roles in various biological processes, such as cell-cell communication and cell-substrate interaction. A sisalyltransferase, ST6GalNAc IV (Neu5Ac-alpha2,3-Gal-beta1,3-GalNAc-alpha2,6-sialyltransferase), catalyzes the formation of alpha2-6-linkages onto GalNAc residues of O-glycosidically linked Ser/Thr of proteins. In this study, we cloned the pig ST6GalNAc IV (pST6GalNAc IV) and investigated its functional characterization. pST6GalNAc IV cDNA has been isolated from pig liver tissues and it contains an entire open reading frame (ORF, 906 bp) coding for 302 amino acid residues. Entire ORF of pST6GalNAc IV containing sialylmotif 'L'-(Large), 'S'-(Small) and '-VS' (Very small) has a high degree of sequence similarity with Homo sapiens (90%), Pan troglodytes (91%) and Mus musculus (87%). Expression of pST6GalNAc IV mRNA in various pig tissues was identified by reverse transcription polymerase chain reaction (RT-PCR) analysis. pST6GalNAc IV mRNA was highly expressed in tongue, muscle and heart, whereas it was not expressed in pancreas. For functional characterization of pST6GalNAc IV gene in pig kidney PK15 cells, we have also established pST6GalNAc IV-transfected PK15 cells, which are stably expressing the pST6GalNAc IV gene. The glycosylation pattern of pST6GalNAc IV-transfected PK15 cells was detected by flow cytometry and immunofluorescence analysis with Maackia amurensis agglutinin (MAA), Maackia amurensis hemagglutinin (MAL II), Sambucus nigra agglutinin (SNA) and peanut agglutinin (PNA) lectins. The specific carbohydrate structures of Neu5Acalpha2-3Galbeta1-3(Neu5Acalpha2-6)GalNAc tetrasaccharide or Neu5Acalpha2-6GalNAc disaccharide recognized by MAL-II and SNA were revealed to be newly synthesized by pST6GalNAc IV. From the results, it was suggested that the pig pST6GalNAc IV gene is capable of synthesizing Neu5Acalpha2-3Galbeta1-3(Neu5Acalpha2-6)GalNAc tetrasaccharide structures on O-glycoproteins.

show abstract

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Molecular characterization of pig α2,3-Gal-β1,3-GalNAc-α2,6-sialyltransferase (pST6GalNAc IV) gene specific for Neu5Acα2-3Galβ1-3GalNAc trisaccharide structure

Song

Jin

et al. 2010

Glycoconj J

View full text Add to dashboard Cite

show abstract

“…Given the ST6Gal-NAc4 high mRNA expression (Fig. 2e) and its restricted substrate specificity, recognizing only sialylated Galβ1-3-GalNAc O-glycans (sialyl T antigen) [18], it is likely that disialyl T antigens are expressed by monocytes and mo-DC.…”

Section: St Involved In α2-6-sialylation Of O-linked Chainsmentioning

confidence: 99%

Surface α2-3- and α2-6-sialylation of human monocytes and derived dendritic cells and its influence on endocytosis

Videira¹,

Amado²,

Crespo³

et al. 2007

Glycoconj J

104

View full text Add to dashboard Cite

Several glycoconjugates are involved in the immune response. Sialic acid is frequently the glycan terminal sugar and it may modulate immune interactions. Dendritic cells (DCs) are antigen-presenting cells with high endocytic capacity and a central role in immune regulation. On this basis, DCs derived from monocytes (mo-DC) are utilised in immunotherapy, though many features are ignored and their use is still limited. We analyzed the surface sialylated glycans expressed during human mo-DC generation. This was monitored by lectin binding and analysis of sialyltransferases (ST) at the mRNA level and by specific enzymatic assays. We showed that alpha 2-3-sialylated O-glycans and alpha 2-6- and alpha 2-3-sialylated N-glycans are present in monocytes and their expression increases during mo-DC differentiation. Three main ST genes are committed with this rearrangement: ST6Gal1 is specifically involved in the augmented alpha 2-6-sialylated N-glycans; ST3Gal1 contributes for the alpha2-3-sialylation of O-glycans, particularly T antigens; and ST3Gal4 may contribute for the increased alpha2-3-sialylated N-glycans. Upon mo-DC maturation, ST6Gal1 and ST3Gal4 are downregulated and ST3Gal1 is altered in a stimulus-dependent manner. We also observed that removing surface sialic acid of immature mo-DC by neuraminidase significantly decreased its endocytic capacity, while it increased in monocytes. Our results indicate the STs expression modulates the increased expression of surface sialylated structures during mo-DC generation, which is probably related with changes in cell mechanisms. The ST downregulation after mo-DC maturation probably results in a decreased sialylation or sialylated glycoconjugates involved in the endocytosis, contributing to the downregulation of one or more antigen-uptake mechanisms specific of mo-DC.

show abstract

Gene‐expression profiles for five key glycosylation genes for galactose‐fed CHO cells expressing recombinant IL‐4/13 cytokine trap

Clark

Griffiths

Bailey

et al. 2005

Biotech & Bioengineering

View full text Add to dashboard Cite

Recombinant protein glycosylation profiles have been shown to affect the in-vivo half-life, and therefore the efficacy and economics, for many therapeutics. While much research has been conducted correlating the effects of various stimuli on recombinant protein glycosylation characteristics, relatively little work has examined glycosylation-related gene-expression profiles. In this study, the effects of galactose feeding on the gene-expression profiles for five key glycosylation-related genes were determined for Chinese hamster ovary cells producing a recombinant IL-4/13 cytokine trap fusion. The genes investigated were sialidase, a putative alpha2,3-sialyltransferase, CMP-sialic acid transporter, beta1,4-galactosyltransferase, and UDP-galactosyltransferase. Additionally, the sialic acid content (sialylation) of the recombinant protein was examined. The peak sialic acid content of the IL-4/13 cytokine trap fusion protein was observed to be similar for the control and galactose-fed cultures. The gene-expression profiles for four of the glycosylation genes were observed to be sensitive to the glucose concentration and not significantly different for the control and galactose-fed cultures prior to glucose depletion. However, the sialidase gene-expression profiles were different for the control and galactose-fed cultures. The sialidase gene-expression profile increased significantly for the galactose-fed cultures prior to glucose depletion, whereas for the control cultures, the sialidase gene-expression profiles did not increase until the late stationary phase. The intracellular sialidase enzyme activity decreased exponentially with time for the control cultures; however, for the galactose-fed cultures, the intracellular sialidase enzyme activity decreased initially and then remained relatively high compared to the control cultures. These results indicate that the galactose feeding may increase the potential for desialylation, which offsets any improvements in the sialylation rate due to increased substrate levels. Thus, galactose feeding is an unnecessary expense for the production of the IL-4/13 cytokine trap fusion protein in a batch process.

show abstract

Cloning, expression and gene organization of a human Neu5Acα2‒3Galβ1‒3GalNAc α2,6-sialyltransferase: hST6GalNAc IV

Cited by 28 publications

References 18 publications

Molecular characterization of pig α2,3-Gal-β1,3-GalNAc-α2,6-sialyltransferase (pST6GalNAc IV) gene specific for Neu5Acα2-3Galβ1-3GalNAc trisaccharide structure

Molecular characterization of pig α2,3-Gal-β1,3-GalNAc-α2,6-sialyltransferase (pST6GalNAc IV) gene specific for Neu5Acα2-3Galβ1-3GalNAc trisaccharide structure

Surface α2-3- and α2-6-sialylation of human monocytes and derived dendritic cells and its influence on endocytosis

Gene‐expression profiles for five key glycosylation genes for galactose‐fed CHO cells expressing recombinant IL‐4/13 cytokine trap

Contact Info

Product

Resources

About