Transfer and expression of a murine UDP-Gal:beta-D-Gal-alpha 1,3-galactosyltransferase gene in transfected Chinese hamster ovary cells. Competition reactions between the alpha 1,3-galactosyltransferase and the endogenous alpha 2,3-sialyltransferase.

Smith, David F.; Larsen, Robert D.; Mattox, Sharon A.; Lowe, John B.; Cummings, Richard D.

doi:10.1016/s0021-9258(19)39314-7

Cited by 118 publications

(16 citation statements)

References 51 publications

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“…These results show that the major isoglobo-GSL isolated from CHO cells transfected with iGb 3 synthase cDNA is iGb 5 . Note that, like parent CHO cells, CHO cells transfected with either murine ␣1,3GalT cDNA (16) or the mutant 199 AVA 201 iGb 3 synthase cDNA only synthesized GlcCer, LacCer, and G M3 in significant amounts, as determined by metabolically labeling with [ 3 H]Gal or [ 3 H]GlcNAc (not shown). A number of additional GSL products from CHO cells transfected with iGb 3 synthase are observed when cells are metabolically labeled with [ 3 H]Gal, but the identity of these products has not yet been established (data not shown).…”

Section: Expression Of Igb 3 Synthase In Cho Cells Leads To the Synthesis Of The Isoglobo-series Gsl-facs Analysis Shows Thatmentioning

confidence: 97%

Expression Cloning of a New Member of the ABO Blood Group Glycosyltransferases, iGb3 Synthase, That Directs the Synthesis of Isoglobo-glycosphingolipids

Keusch¹,

Manzella²,

Nyame³

et al. 2000

Journal of Biological Chemistry

108

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The large array of different glycolipids described in mammalian tissues is a reflection, in part, of diverse glycosyltransferase expression. Herein, we describe the cloning of a UDP-galactose: beta-d-galactosyl-1,4-glucosylceramide alpha-1, 3-galactosyltransferase (iGb(3) synthase) from a rat placental cDNA expression library. iGb(3) synthase acts on lactosylceramide, LacCer (Galbeta1,4Glcbeta1Cer) to form iGb(3) (Galalpha1,3Galbeta1, 4Glcbeta1Cer) initiating the synthesis of the isoglobo-series of glycosphingolipids. The isolated cDNA encoded a predicted protein of 339 amino acids, which shows extensive homology (40-50% identity) to members of the ABO gene family that includes: murine alpha1, 3-galactosyltransferase, Forssman (Gb(5)) synthase, and the ABO glycosyltransferases. In contrast to the murine alpha1, 3-galactosyltransferase, iGb(3) synthase preferentially modifies glycolipids over glycoprotein substrates. Reverse transcriptase-polymerase chain reaction revealed a widespread tissue distribution of iGb(3) synthase RNA expression, with high levels observed in spleen, thymus, and skeletal muscle. As an indirect consequence of the expression cloning strategy used, we have been able to identify several potential glycolipid biosynthetic pathways where iGb(3) functions, including the globo- and isoglobo-series of glycolipids.

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Section: Expression Of Igb 3 Synthase In Cho Cells Leads To the Synthesis Of The Isoglobo-series Gsl-facs Analysis Shows Thatmentioning

confidence: 97%

Expression Cloning of a New Member of the ABO Blood Group Glycosyltransferases, iGb3 Synthase, That Directs the Synthesis of Isoglobo-glycosphingolipids

Keusch¹,

Manzella²,

Nyame³

et al. 2000

Journal of Biological Chemistry

108

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“…CHO and Lec8 CHO cells (Stanley, 1985 ;Deutscher and Hirschberg, 1986) were grown in a-modified Eagles' medium (a-MEM) supplemented with 10% FCS. NeoLewis CHO cells, initially designated as CHO-FT cells , and Clone 3 CHO cells (Smith et al ., 1990) were cultured in a-MEM/10% FCS supplemented with G-418 (Gibco Laboratories, Grand Island, NY) at 400 pg/ml of active drug. F9 teratocarcinoma cells were cul-The Journal of Cell Biology, Volume 115, 1991 tured and induced to differentiate with retinoic acid (RA/F9) as described previously (Cummings and Mattox, 1988) .…”

Section: Cellsmentioning

confidence: 99%

The selectin GMP-140 binds to sialylated, fucosylated lactosaminoglycans on both myeloid and nonmyeloid cells.

Zhou

Moore

Smith

et al. 1991

The Journal of Cell Biology

Self Cite

172

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Abstract. is an inducible receptor for myeloid leukocytes on activated platelets and endothelium . Like other selectins, GMP140 recognizes specific oligosaccharide ligands . However, prior data on the nature of these ligands are contradictory. We investigated the structural features required for ligand interaction with GMP-140 using purified GMP-140, cells naturally expressing specific oligosaccharides, and cells expressing cloned glycosyltransferases . Like the related selectin endothelial leukocyte adhesion molecule-1 (ELAM-1), GMP-140 recognizes a(2-3)sialylated, a(1-3)fucosylated lactosaminoglycans on both myeloid and nonmyeloid cells,

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“…Host cells transfected with several copies of the α1,3GT gene (GGTA1) —stable transfection of host cells with several copies of the α1,3GT gene ( GGTA1 ) is likely to result in increased concentration of α1,3GT in the trans-Golgi to levels that are much higher than the natural concentration of the enzyme in non-primate mammalian cells. Such stable transfection will increase the probability of capping viral N-glycans of the complex type with α-gal epitopes, rather than with sialic acid ( Smith et al, 1990 ). In host cells originating in Old-World monkeys, such as Vero cells (African green monkey cells) and in human cells, production of α1,3GT by several copies of the α1,3GT transgene is likely to ensure synthesis of multiple α-gal epitopes on viral glycans, as well.…”

Section: Immunological Processes Associated With Anti-gal/α-gal Epitope Interactions Which May Be Harnessed For α-Gal Therapiesmentioning

confidence: 99%

“…It is synthesized by the glycosylation enzyme α1,3galactosyltransferase (α1,3GT) ( Galili et al, 1988a ; Basu and Basu, 1973 ; Betteridge and Watkins, 1983 ; Blake and Goldstein, 1981 ; Blanken and Van den Eijnden, 1985 ). This enzyme is active in the trans-Golgi apparatus ( Smith et al, 1990 ), linking galactose to N-acetyllactosaminyl groups (Galβ1-4GlcNAc-R) by using UDP-Gal as the sugar donor ( Figure 2 ) and forming the trisaccharide Galα1-3Galβ1-4GlcNAc-R on various glycans (right glycan in Figure 2 ). In the trans-Golgi, α1,3GT competes mostly with sialyltransferases which cap nascent glycans with sialic acid (left glycan in Figure 2 ) ( Smith et al, 1990 ).…”

Section: Introductionmentioning

confidence: 99%

“…This enzyme is active in the trans-Golgi apparatus ( Smith et al, 1990 ), linking galactose to N-acetyllactosaminyl groups (Galβ1-4GlcNAc-R) by using UDP-Gal as the sugar donor ( Figure 2 ) and forming the trisaccharide Galα1-3Galβ1-4GlcNAc-R on various glycans (right glycan in Figure 2 ). In the trans-Golgi, α1,3GT competes mostly with sialyltransferases which cap nascent glycans with sialic acid (left glycan in Figure 2 ) ( Smith et al, 1990 ). The number of α-gal epitopes per cell differs from one tissue to the other and in various mammalian species and depends on the activity of α1,3GT vs. that of competing sialyltransferases or other capping transferases within the trans-Golgi.…”

Section: Introductionmentioning

confidence: 99%

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Biosynthesis of α-Gal Epitopes (Galα1-3Galβ1-4GlcNAc-R) and Their Unique Potential in Future α-Gal Therapies

Galili

2021

Front. Mol. Biosci.

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The α-gal epitope is a carbohydrate antigen which appeared early in mammalian evolution and is synthesized in large amounts by the glycosylation enzyme α1,3galactosyltransferase (α1,3GT) in non-primate mammals, lemurs, and New-World monkeys. Ancestral Old-World monkeys and apes synthesizing α-gal epitopes underwent complete extinction 20–30 million years ago, and their mutated progeny lacking α-gal epitopes survived. Humans, apes, and Old-World monkeys which evolved from the surviving progeny lack α-gal epitopes and produce the natural anti-Gal antibody which binds specifically to α-gal epitopes. Because of this reciprocal distribution of the α-gal epitope and anti-Gal in mammals, transplantation of organs from non-primate mammals (e.g., pig xenografts) into Old-World monkeys or humans results in hyperacute rejection following anti-Gal binding to α-gal epitopes on xenograft cells. The in vivo immunocomplexing between anti-Gal and α-gal epitopes on molecules, pathogens, cells, or nanoparticles may be harnessed for development of novel immunotherapies (referred to as “α-gal therapies”) in various clinical settings because such immune complexes induce several beneficial immune processes. These immune processes include localized activation of the complement system which can destroy pathogens and generate chemotactic peptides that recruit antigen-presenting cells (APCs) such as macrophages and dendritic cells, targeting of antigens presenting α-gal epitopes for extensive uptake by APCs, and activation of recruited macrophages into pro-reparative macrophages. Some of the suggested α-gal therapies associated with these immune processes are as follows: 1. Increasing efficacy of enveloped-virus vaccines by synthesizing α-gal epitopes on vaccinating inactivated viruses, thereby targeting them for extensive uptake by APCs. 2. Conversion of autologous tumors into antitumor vaccines by expression of α-gal epitopes on tumor cell membranes. 3. Accelerating healing of external and internal injuries by α-gal nanoparticles which decrease the healing time and diminish scar formation. 4. Increasing anti-Gal–mediated protection against zoonotic viruses presenting α-gal epitopes and against protozoa, such as Trypanosoma, Leishmania, and Plasmodium, by vaccination for elevating production of the anti-Gal antibody. The efficacy and safety of these therapies were demonstrated in transgenic mice and pigs lacking α-gal epitopes and producing anti-Gal, raising the possibility that these α-gal therapies may be considered for further evaluation in clinical trials.

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Transfer and expression of a murine UDP-Gal:beta-D-Gal-alpha 1,3-galactosyltransferase gene in transfected Chinese hamster ovary cells. Competition reactions between the alpha 1,3-galactosyltransferase and the endogenous alpha 2,3-sialyltransferase.

Cited by 118 publications

References 51 publications

Expression Cloning of a New Member of the ABO Blood Group Glycosyltransferases, iGb3 Synthase, That Directs the Synthesis of Isoglobo-glycosphingolipids

Expression Cloning of a New Member of the ABO Blood Group Glycosyltransferases, iGb3 Synthase, That Directs the Synthesis of Isoglobo-glycosphingolipids

The selectin GMP-140 binds to sialylated, fucosylated lactosaminoglycans on both myeloid and nonmyeloid cells.

Biosynthesis of α-Gal Epitopes (Galα1-3Galβ1-4GlcNAc-R) and Their Unique Potential in Future α-Gal Therapies

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