Stable Expression of Blood Group H Determinants and GDP-L-fucose: β-D-galactoside 2-α-L-Fucosyltransferase in Mouse Cells After Transfection with Human DNA

Эрнст, Л. К.; Rajan, Valanila P.; Larsen, Robert D.; Ruff, Melissa M.; Lowe, John B.

doi:10.1016/s0021-9258(18)94086-x

Cited by 91 publications

(5 citation statements)

References 66 publications

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“…COS-7 cells, harvested 72 h after transfection, were stained with monoclonal antibodies diluted in staining media, using methods described previously [19,20]. Anti-A, -B and -H antibodies were used at 10 µg\ml.…”

Section: Flow-cytometry Analysismentioning

confidence: 99%

“…Murine genomic DNA was prepared from C57BL\6 mouse liver, subjected to restriction digestion, fractionated by electrophoresis on 1.2 % (w\v) agarose gels and transferred to Hybond-N membranes (Amersham Life Sciences, Arlington Heights, IL, U.S.A.) as described previously [20,25]. Blots were probed with either the 1.2 kb HinfI fragment of the human FUT1 [5,6], or a murine FUT1 gene-specific probe (probe A) generated by PCR amplification of cloned template DNA using the following primers : forward primer TTCCTGTCCTGAGCAGTCCTCC-TCACTCTCTGGGA ; reverse primer GAATTCCCTACGAA-TCTGTTCCCGAAGATGATG (the locations are indicated on Figure 1B).…”

Section: Southern Blottingmentioning

confidence: 99%

“…Poly(A) + RNA samples were prepared by oligo(dT) column chromatography, subjected to gel electrophoresis and transferred to Hybond-N as described previously [16]. Pre-hybridization and hybridization were performed at 42 mC in 50% (w\v) formamide, 5iSSC, 1iPE [0.25 M Tris\HCl, pH 7.5\0.025 M EDTA\0.5 % (w\v) sodium pyrophosphate\5 % (w\v) SDS\1 % (w\v) poly(vinylpyrolidone)\1 % (w\v) Ficoll\1 % (w\v) BSA] and 150 mg\ml of sheared salmon sperm DNA [20]. Blots were hybridized for 24-48 h at 42 mC using probe B labelled with [α-$#P]dCTP to a specific radioactivity of 1i10*-2i10* c.p.m.\µg DNA using the Rediprime DNA labelling kit (Amersham).…”

Section: Northern-blot Analysismentioning

confidence: 99%

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Molecular cloning, chromosomal assignment and tissue-specific expression of a murine α(1,2)fucosyltransferase expressed in thymic and epididymal epithelial cells

Hiraiwa

Lowe

1997

Biochemical Journal

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Terminal Fucalpha(1-2)Galbeta epitopes have been proposed to play significant roles in cell-cell interactions in development, cell adhesion, and malignant transformation. To begin to investigate the regulation and function of alpha(1-2)fucosylated epitopes in an animal model, we have isolated and characterized a mouse genomic DNA segment encoding a protein orthologous to the human H blood group locus alpha(1,2)fucosyltransferase (FUT1). This segment maintains an open reading frame encoding 376 amino acids sharing 75% sequence identity with the enzyme encoded by human FUT1, and 55% sequence identity with the enzyme encoded by the human Secretor blood group locus (FUT2). Expression of the open reading frame in COS-7 cells yields an alpha(1,2)fucosyltransferase activity with a Km of 7.6 mM for phenyl-beta-d-galactoside. Southern blotting and interspecific backcross analyses indicate that this murine locus represents a single copy sequence mapping to a novel locus 2.1 centimorgans from the Klk1 locus, in a region of homology between mouse chromosome 7 and the human FUT1 locus on the long arm of chromosome 19. Mouse FUT1 yields a 2.8 kb mRNA transcript identifiable in many organs, including thymus, lung, stomach, pancreas, small intestine, colon, uterus and epidiymis. Hybridization analyses in situ localize expression of FUT1 transcripts to thymic medullary and epididymal epithelial cells, implying that this gene determines the expression of cell surface Fucalpha(1-2)Galbeta epitopes in these tissues.

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Section: Flow-cytometry Analysismentioning

confidence: 99%

Section: Southern Blottingmentioning

confidence: 99%

Section: Northern-blot Analysismentioning

confidence: 99%

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Molecular cloning, chromosomal assignment and tissue-specific expression of a murine α(1,2)fucosyltransferase expressed in thymic and epididymal epithelial cells

Hiraiwa

Lowe

1997

Biochemical Journal

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“…The radiolabeled cell monolayers were harvested in parallel with the nonlabeled monolayers and pooled with the nonlabeled cells. The cells were then pelleted and subjected to a series of solvent extractions before pronase digestion, as described previously (18)(19)(20). Briefly, the cell pellets were extract three times with chloroform/methanol (2:1), once with methanol, and then twice with water.…”

Section: Preparation Of Gifopeptidesmentioning

confidence: 99%

Protective effects of sialylated oligosaccharides in immune complex-induced acute lung injury.

Mulligan

Lowe

Larsen

et al. 1993

The Journal of Experimental Medicine

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150

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SummaryUsing sialyl Lewis x (SLX) oligosaccharides derived from fucosyl transferase-expressing cells or generated synthetically, the ability of these compounds to protect against acute lung damage after deposition of immunoglobulin (Ig)G or IgA immune complexes has been determined. The synthetic compounds were tetra-and pentasaccharide derivates of SLX as well as the nonfucosylated forms of SLX as controls. In the IgG immune complex modal of lung injury, which is E-selectin dependent, SLX preparations provided dose-dependent protective effects, as assessed by changes in lung vascular permeability and hemorrhage. Protective effects were associated with diminished tissue accumulation of neutrophils in lungs (as assessed by myeloperoxidase). Morphological assessment revealed reduced physical contact of neutrophils with the pulmonary vascular endothelium and reduced tissue accumulation of neutrophils. In the model of IgA immune complex-induced lung injury, which does not involve participation of neutrophils and is independent of the requirement for E-selectin, SLX preparations were not protective. These data suggest that, in neutrophil-mediated and E-sdectin-dependent lung injury, SLX preparations provide significant, protective effects against inflammatory vascular injury. The ability to achieve antiinflammatory outcomes in vivo with appropriate oligosaccharides suggests a new approach to the blocking of acute inflammatory responses.

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“…These findings are consistent with the notion that the HB-6 Ag is a carbohydrate structure that is expressed on cell surface glycoproteins and glycolipids and one that contains an a2,6-linked sialic acid residue as a critical component of the Ag . The isolation of a cDNA which encodes a glycosyltransferase by the selection of transfected cells which express the product of the glycosyltransferase at the cell surface is not without precedent as the cDNA for several terminal glycosyltransferases have been isolated by expression cloning (13,30,31). Most recently, a cDNA encoding an a-1,3-fucosyltransferase, which acts in the synthesis of a carbohydrate moiety that constitutes the ELAM-1 ligand, was isolated by expression cloning (19) .…”

Section: Discussionmentioning

confidence: 99%

The HB-6, CDw75, and CD76 differentiation antigens are unique cell-surface carbohydrate determinants generated by the beta-galactoside alpha 2,6-sialyltransferase.

Bast

Zhou

Freeman

et al. 1992

The Journal of Cell Biology

113

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Abstract. Expression of the ß-galactoside a2,6-sialyltransferase (a2,6-ST) was shown to regulate the generation of multiple cell-surface differentiation antigens (Ags) that may be necessary for lymphocyte function . A new mAb was produced, termed HB-6, that was shown to identify a novel neuraminidasesensitive cell-surface Ag expressed by subpopulations of human lymphocytes and erythrocytes . In attempting to isolate a cDNA encoding the HB-6 antigen by expression cloning, a cDNA encoding the a2,6-ST (EC 2 .4.99.1) was obtained. Since expression of the a2,6-ST protein was shown to be limited to the Golgi apparatus, the cell-surface HB-6 Ag was demonstrated to be the product of c12,6-ST activity. Interestingly, a2,6-T HE identification of leukocyte cell-surface structures, delineation of their primary amino acid sequences, and elucidation of their potential functions has been a major focus in the field of immunology during the last decade. Each cell-surface protein has been found to have a unique primary sequence that presumably directs function of the molecule. However, these proteins may also serve as scaffolds for the presentation of cell-surface carbohydrates . It is likely that cell-surface carbohydrates are also functional moieties with a broad potential for diversity not only in the proteins upon which they may appear, but also in the complexity ofcarbohydrate side chains possible (46,49,51) . In this study, we have examined some of the potential diversity in three leukocyte cell surface antigens (Ag)', HB-6, CDw75, and CD76.

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Stable Expression of Blood Group H Determinants and GDP-L-fucose: β-D-galactoside 2-α-L-Fucosyltransferase in Mouse Cells After Transfection with Human DNA

Cited by 91 publications

References 66 publications

Molecular cloning, chromosomal assignment and tissue-specific expression of a murine α(1,2)fucosyltransferase expressed in thymic and epididymal epithelial cells

Molecular cloning, chromosomal assignment and tissue-specific expression of a murine α(1,2)fucosyltransferase expressed in thymic and epididymal epithelial cells

Protective effects of sialylated oligosaccharides in immune complex-induced acute lung injury.

The HB-6, CDw75, and CD76 differentiation antigens are unique cell-surface carbohydrate determinants generated by the beta-galactoside alpha 2,6-sialyltransferase.

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