Identification and Characterization of UDP-GalNAc: NeuAcα2–3Galβ1–4Glc(NAc) β1–4(GalNAc to Gal)N-Acetylgalacto saminyltransferase in Human Blood Plasma1

Takeya, Akira; Hosomi, Osamu; Kogure, Tadahisa

doi:10.1093/oxfordjournals.jbchem.a121898

Cited by 31 publications

(15 citation statements)

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“…The B4GALNT2 enzyme, isolated from various human and animal tissues, catalyzes the synthesis of the SDa blood group antigen by the addition of beta 1,4‐linked GalNAc to an alpha 2,3 sialic acid‐modified N‐acetyl lactosamine acceptor oligosaccharide. The B4GALNT2 enzyme preferentially utilizes N‐acetylneuraminic acid (Neu5Ac) α2,3 galactose β1,4 glucose as an acceptor molecule and does not react with Neu5Ac α2,6‐modified lactose.…”

Section: The Sda Human Blood Groupmentioning

confidence: 99%

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B4GALNT2 and xenotransplantation: A newly appreciated xenogeneic antigen

Byrne

Ahmad-Villiers

et al. 2018

Xenotransplantation

100

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Analysis of non-Gal antibody induced after pig-to-baboon cardiac xenotransplantation identified the glycan produced by porcine beta-1,4-N-acetyl-galactosaminyltransferase 2 (B4GALNT2) as an immunogenic xenotransplantation antigen. The porcine B4GALNT2 enzyme is homologous to the human enzyme, which synthesizes the human SDa blood group antigen. Most humans produce low levels of anti-SDa IgM which polyagglutinates red blood cells from rare individuals with high levels of SDa expression. The SDa glycan is also present on GM2 gangliosides. Clinical GM2 vaccination studies for melanoma patients suggest that a human antibody response to SDa can be induced. Expression of porcine B4GALNT2 in human HEK293 cells results in increased binding of anti-SDa antibody, and increased binding of Dolichos biflorus agglutinin (DBA) a lectin commonly used to detect SDa. In pigs, B4GALNT2 is expressed by vascular endothelial cells and endothelial cells from a wide variety of pig backgrounds stain with DBA suggesting that porcine vascular expression of B4GALNT2 is not polymorphic. Mutations in B4GALNT2 have been engineered in mice and pigs. In both species, the B4GALNT2-KO animals are apparently normal and no longer show evidence of SDa antigen expression. Pig tissues with a mutation in B4GALNT2, added to a background of alpha-1,3-galactosyltransferase deficient (GGTA1-KO) and cytidine monophosphate-N-acetylneuraminic acid hydroxylase deficient (CMAH-KO), show reduced antibody binding, confirming the presence of B4GALNT2 dependent antibodies in both humans and nonhuman primates. Preclinical xenotransplantation using B4GALNT2 deficient donors has recently been reported. Elimination of this source of immunogenic pig antigen should minimize acute injury by preformed anti-pig antibody and eliminate an induced clinical immune response to this newly appreciated xenotransplantation antigen.

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Section: The Sda Human Blood Groupmentioning

confidence: 99%

“…The B4GALNT2 enzyme, isolated from various human [34][35][36] and animal tissues, [37][38][39][40] catalyzes the synthesis of the SDa blood group antigen by the addition of beta 1,4-linked GalNAc to an alpha 2,3 sialic acid-modified N-acetyl lactosamine acceptor oligosaccharide.…”

Section: The S Da H Uman B Lood G Roupmentioning

confidence: 99%

B4GALNT2 and xenotransplantation: A newly appreciated xenogeneic antigen

Byrne

Ahmad-Villiers

et al. 2018

Xenotransplantation

100

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“…Sialylated products were resolved on an HPLC system equipped with an ion-exchange column, PALPAK Type N (6.4 mm × 25 cm) (Takara Bio, Inc.). Chromatography was performed at 40 • C with a single solvent system of CH 3 [11,12,32,33] with slight modifications. The reaction mixture had the following final concentration in a total volume of 20 µl: 0.5 mg/ml glycoproteins, 1 mM CMP-Neu5Ac, Triton CF-54 at the indicated concentrations, 1 mM MnCl 2 , 25 mM sodium cacodylate-HCl buffer, pH 6.5, and enzyme fractions.…”

Section: Sialyltransferase Assaymentioning

confidence: 99%

Purification and characterization of a soluble recombinant human ST6Gal I functionally expressed in Escherichia coli

et al. 2005

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A soluble and active form of recombinant human ST6Gal I was expressed in Escherichia coli. The gene encoding the soluble form of ST6Gal I lacking the membrane and cytosolic regions was introduced into a bacterial expression vector, pMAL-p2X, fused in frame with a maltose-binding protein (MBP) tag. Low-temperature cultivation at 13 degrees C during IPTG-induction significantly improved both solubility and MBP-tagging of the recombinant enzyme expressed in bacteria. The supernatant prepared by disruption of the cells demonstrated sialic acid transfer activity to both an oligosaccharide and a glycoprotein, asialofetuin, indicating that the enzyme expressed in bacteria is soluble and active. The MBP-tagged enzyme was efficiently purified by a combination of cation-exchange column and amylase-conjugated agarose column chromatography. The purified recombinant enzyme exerted enzymatic activity even in the absence of detergents in the reaction mixture. Acceptor substrate specificity of the enzyme was marginally different from that of rat liver ST6Gal I. These observations suggest that membrane and cytosolic regions of ST6Gal I may affect the properties of the enzyme. The purified recombinant enzyme was applied to convert desialylated fetuin to resialylated fetuin. Lectin blotting demonstrated that resialylated fetuin possesses a single Neu5Ac alpha 2-6 residue. The resialylated fetuin efficiently blocked hemagglutination induced by influenza virus strain A/Memphis/1/71 (H3N2), indicating that resialylated carbohydrate chains on the protein are so active as to competitively inhibit virus-receptor interaction. In conclusion, soluble recombinant ST6Gal I obtained using our bacterial expression system is a valuable tool to investigate the molecular mechanisms of biological and pathological interactions mediated via carbohydrates.

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“…The Sd a β1,4GalNAc transferase activity has been described essentially in human intestine [11], colon [4] and kidney [12]. In addition, this enzyme activity has been detected in different body fluids, such as blood plasma [13] and urine [14]. In both colon and kidney, this enzyme appears to be onco-developmentally regulated.…”

Section: Introductionmentioning

confidence: 99%

Molecular cloning, gene organization and expression of the human UDP-GalNAc:Neu5Acalpha2-3Galbeta-R beta1,4-N-acetylgalactosaminyltransferase responsible for the biosynthesis of the blood group Sda/Cad antigen: evidence for an unusual extended cytoplasmic domain

Montiel

Krzewinski-Recchi

Delannoy

et al. 2003

Biochemical Journal

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The nucleotide sequence of the short and long transcripts of beta1,4- N -acetylgalactosaminyltransferase have been submitted to the DDBJ, EMBL, GenBank(R) and GSDB Nucleotide Sequence Databases under accession nos AJ517770 and AJ517771 respectively. The human Sd(a) antigen is formed through the addition of an N -acetylgalactosamine residue via a beta1,4-linkage to a sub-terminal galactose residue substituted with an alpha2,3-linked sialic acid residue. We have taken advantage of the previously cloned mouse cDNA sequence of the UDP-GalNAc:Neu5Acalpha2-3Galbeta-R beta1,4- N -acetylgalactosaminyltransferase (Sd(a) beta1,4GalNAc transferase) to screen the human EST and genomic databases and to identify the corresponding human gene. The sequence spans over 35 kb of genomic DNA on chromosome 17 and comprises at least 12 exons. As judged by reverse transcription PCR, the human gene is expressed widely since it is detected in various amounts in almost all cell types studied. Northern blot analysis indicated that five Sd(a) beta1,4GalNAc transferase transcripts of 8.8, 6.1, 4.7, 3.8 and 1.65 kb were highly expressed in colon and to a lesser extent in kidney, stomach, ileum and rectum. The complete coding nucleotide sequence was amplified from Caco-2 cells. Interestingly, the alternative use of two first exons, named E1(S) and E1(L), leads to the production of two transcripts. These nucleotide sequences give rise potentially to two proteins of 506 and 566 amino acid residues, identical in their sequence with the exception of their cytoplasmic tail. The short form is highly similar (74% identity) to the mouse enzyme whereas the long form shows an unusual long cytoplasmic tail of 66 amino acid residues that is as yet not described for any other mammalian glycosyltransferase. Upon transient transfection in Cos-7 cells of the common catalytic domain, a soluble form of the protein was obtained, which catalysed the transfer of GalNAc residues to alpha2,3-sialylated acceptor substrates, to form the GalNAcbeta1-4[Neu5Acalpha2-3]Galbeta1-R trisaccharide common to both Sd(a) and Cad antigens.

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Identification and Characterization of UDP-GalNAc: NeuAcα2–3Galβ1–4Glc(NAc) β1–4(GalNAc to Gal)N-Acetylgalacto saminyltransferase in Human Blood Plasma1

Cited by 31 publications

References 0 publications

B4GALNT2 and xenotransplantation: A newly appreciated xenogeneic antigen

B4GALNT2 and xenotransplantation: A newly appreciated xenogeneic antigen

Purification and characterization of a soluble recombinant human ST6Gal I functionally expressed in Escherichia coli

Molecular cloning, gene organization and expression of the human UDP-GalNAc:Neu5Acalpha2-3Galbeta-R beta1,4-N-acetylgalactosaminyltransferase responsible for the biosynthesis of the blood group Sda/Cad antigen: evidence for an unusual extended cytoplasmic domain

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