Glycoprotein Alpha 1,3-Galactosyltransferase 1, Pseudogene (GGTA1P)

Reference work entry


UDPgalactose:β-d-galactosyl-1,4-N-acetyl-d-glucosaminide α1,3 galactosyltransferase, α1,3GT catalyzes the synthesis of the α-Gal epitope, one of the most common carbohydrate structures found in mammalian tissues (Galili and Tanemura 1999). This gene is present in the DNA of humans and Old World monkeys, but is not transcribed by evolutionary inactivation (Joziasse et al. 1992).


World Monkey Sialyl LewisX Clinical Xenotransplantation Acceptor Substrate Specificity Nuclear Transplantation Technique 


UDPgalactose:β-d-galactosyl-1,4-N-acetyl-d-glucosaminide α1,3 galactosyltransferase, α1,3GT catalyzes the synthesis of the α-Gal epitope, one of the most common carbohydrate structures found in mammalian tissues (Galili and Tanemura 1999). This gene is present in the DNA of humans and Old World monkeys, but is not transcribed by evolutionary inactivation (Joziasse et al. 1992).

α1,3GT activity was first detected in mouse Ehrlich ascites tumor cells (Blake and Goldstein 1981). The cDNA encoding α1,3GT was cloned from both the bovine and mouse sources (Joziasse et al. 1989; Larsen et al. 1989). The enzyme accepts various disaccharides as substrates in addition to LacNAc, such as lactose and Gal derivatives, albeit with lower specific activities (Shah et al. 2000). It shares its oligosaccharide specificity with α1,2FucT,α1,3FucT,α2,3NeuAcT and α2,6NeuAcT (Sepp et al. 1997). The enzyme different from the blood group B transferase, another α1,3GT that has a different acceptor substrate specificity.

Mouse and pig α1,3GT have been knocked out. α1,3GT-KO pigs were produced by nuclear transfer and have contributed to the success of clinical xenotransplantation (Dai et al. 2002; Takahagi et al.2005).


NC-IUBMB enzyme classification: E.C.

Glycoprotein alpha 1,3-galactosyltransferase 1, pseudogene (GGTA1P)






Homo sapiens












Mus musculus
















Rattus norvegicus












Sus scrofa







Bos taurus






Canis lupus familiaris





Ovis aries







Felis catus





Callithrix jacchus





Cricetulus griseus





Oryctolagus cuniculus




Alouatta caraya





Loris tardigradus





Tarsius syrichta





Cebus capucinus





Pan troglodytes





Pan paniscus



Gorilla gorilla



Pongo pygmaeus



Macaca mulatta



Chlorocebus aethiops



Erythrocebus patas



Ateles geoffroyi



Saimiri sciureus



Alouatta caraya



Mustela putorius furo




Name and History

The glycosylation enzyme α1,3 galactosyltransferase (α1,3GT; UDPgalactose:β-d-galactosyl-1,4-N-acetyl-d-glucosaminide α1,3 galactosyltransferase) catalyzes the synthesis of the α-Gal epitope. The enzyme α 1,3 GT catalyzes the following reaction:
$$ \begin{array}{l}\mathrm{Gal}\ \upbeta 1,4\mathrm{GlcNAc}\hbox{-} \mathrm{R}+\mathrm{UDP}\hbox{-} \mathrm{Gal}\ \left(\upalpha 1,3\mathrm{GT}\right)\\ {}\kern4em \hbox{-} \hbox{-} \hbox{-} \hbox{-} \mathrm{Gal}\upalpha 1,3\mathrm{Gal}\ \upbeta 1,4\mathrm{GlcNAc}\hbox{-} \mathrm{R}+\mathrm{UDP}\end{array} $$

The α-Gal epitope is one of the most common carbohydrate structures found in mammalian tissues (Galili and Tanemura 1999). The epitope was first identified as a major component of rabbit erythrocyte glycolipids, ceramide pentahexoside (CPH), and was reported to be an internal type 1 chain (Eto et al. 1968) and later corrected to be a linear type 2 chain oligosaccharide (Stellner et al. 1973). The α-Gal epitope, with carbohydrate chains of various lengths, was subsequently shown to be present on rabbit red cell glycolipids (Homma et al.1981; Egge et al. 1985).

The α1,3GT enzyme is found in the trans-Golgi network where it catalyzes the production of α-Gal epitopes using N-acetyl lactosaminyl residues as the sugar acceptor and UDP-Gal as the sugar donor.

α1,3GT activity was first detected in mouse Ehrlich ascites tumor cells (Blake and Goldstein 1981). The cDNA encoding α1,3GT was cloned from both bovine and mouse sources (Joziasse et al. 1989; Larsen et al. 1989). The enzyme is different from the blood group B transferase, another α1,3GT that has a different acceptor substrate specificity.

This gene is present in the DNA of humans and Old World monkeys, but is not transcribed by them, suggesting the complete lack of α1,3GT mRNA (Joziasse et al. 1989). These findings suggest that the evolutionary inactivation of the α1,3GT gene in ancestral primates was likely associated with mutations in the regulatory domain(s) of this gene (e.g., mutations in the promoter/enhancer domains) (Joziasse et al. 1992). Frameshift mutations were found in the open reading frame of the human α1,3GT pseudogene, which results in premature stop codons (Larsen et al. 1990). Shortly after the above study, similar frameshift mutations were found in apes, but not in Old World monkeys (Galili and Swanson 1991). Thus, the lack of α1,3GT activity in humans appears to be the result of both regulatory and structural mutations within the α1,3GT gene.


The structure of the molecule was reported by several groups. Minimum activity was observed in the 94 amino acids from the C-terminal of the enzyme of the marmoset (Henion et al. 1994), and the importance of E317 was mentioned for covalent binding to galactose, as evidence by X-ray crystal structural analysis. The crystal structure of the catalytic domain of substrate-free bovine α1,3GT, solved and refined to 2.3 A resolution, has a globular shape with an alpha/beta fold containing a narrow cleft on one face and shares a UDP-binding domain (UBD) with the recently solved inverting glycosyltransferases. The substrate-bound complex, solved and refined to 2.5 A, allows the description of residues interacting directly with UDP-galactose (Gastinel et al. 2001). However, E317 may not be a catalytic nucleophile (Molina et al. 2007). On the other hand, His271, Trp356, and the negative charge of Asp316 also appear to be important for catalytic activity (Lazarus et al. 2002; Zhang et al. 2003, 2004; Tumbale et al. 2008).

Enzyme Activity Assay and Substrate Specificity

The assay for the enzyme is usually performed using UDP-[3H] or -[14C]galactose (Zhang et al. 2001). As an alternative to the use of radioactive materials, the sample cell can be sonicated and lysed in PBS. The acceptor substrate, pyridylaminated lacto-N-neotetraose (LNnT-PA) (Galβ1-4GlcNAcβ1-3Galβ1-4Glc-PA) at a final concentration of 10 μM, is then employed in the activity assays. Lacto-N-neotetraose is prepared and pyridylaminated. The activity is assayed in a reaction mixture containing 10 μM HEPES, pH 7.2, 20 mM UDP-galactose, 10 mM MnCl2, 33 mM NaCl, and 3 mM KCl. A 10 μl of a 50 μM substrate and 15 μl of cell lysate are added to this mixture, which is then incubated at 37 °C for 3 h. The enzyme reactions are quenched by boiling for 5 min. The samples are then centrifuged at 12,000 × g for 5 min, and an aliquot of each supernatant is subjected to HPLC analysis, using a TSK-gel ODS-80TM column (4.6 × 250 mm). The reaction products are eluted with 20 mM acetate buffer, at pH 4.0 containing n-butanol at a flow rate of 1.0 ml/min at 55 °C and are monitored with a fluorescence spectrophotometer using excitation and emission wavelengths of 320 and 400 nm, respectively. The specific activity of the enzyme is expressed as moles of product produced per hour of incubation per mg of protein. Protein concentrations are determined by means of a BCA protein assay kit, using bovine serum albumin as a standard.

In addition to LacNAc, which is the natural substrate, the enzyme is capable of accepting various other disaccharides as substrates such as lactose and Gal derivatives, β-O-methylgalactose and β-d-thiogalactopyranoside, albeit the specific activities are lower. There is an absolute requirement for Gal to be at the nonreducing end of the acceptor molecule that must be β1-4 linked to a second residue that can be more diverse in structure. The second monosaccharide is critical for holding the acceptor molecule in place (Shah et al. 2000).

It shares its oligosaccharide specificity with α1,2 FucT, leading to the type 2 H epitope, α1,3FucT to synthesize the LewisX epitope, α2,3 NeuAcT to produce the precursor of sialyl LewisX, α2,6NeuAcT, ultimately leading to the expression of the CDw75, CD76, and HB-6 carbohydrate epitopes (Sepp et al. 1997). Although α1,3GT and α2,6NeuAcT potentially compete for common LacNAc acceptor sites on protein-linked glycans, they may in fact cooperate in the case of branched glycans because they prefer different branches on such substrates (α2,6NeuAcT; Manα1,3Man branch; α1,3GT: Manα1,6Man branch) (Van den Eijnden 2000).

In addition to the OH group at C-3 of the Gal in Galβ1-4GlcNAc, which is the site at which α-Gal is introduced, the OH group at C-4 of the Gal is required for activity. Deoxygeneration of this OH yields an inactive acceptor. OH groups that may be deoxygenated but may not be substituted without loss of activity are these at the C-2 and C-6 positions of Gal and the C-3 position of GlcNAc (Sujino et al. 1997).


α1,3GT can be obtained from calf thymus in a highly active form. After solubilization in Triton X-100, it is purified by repeated affinity chromatography on a column of UDP-hexanolamine-Sepharose (Blanken and Van den Eijnden 1985). Separation from β4GalT can be achieved by chromatography on a column of α-lactalbumin to which α1,3GT does not bind. In general, a recombinant form of the enzyme can be prepared by expression in E. coli (unglycosylated), insect cells (high mannose rich), mammalian cells, etc. The enzymatically active recombinant α1,3GT can be prepared by expression in E.coli (Zhang et al. 2001), and this method is relatively popular. On the other hand, the introduction of an insect signal peptide in the expression construct permits the enzymatically active α1,3GT to be secreted. This recombinant enzyme form has been used in an efficient one-pot synthesis of the Galα1-3Galβ1-4GlcNAc sequence (Joziasse et al. 1990; Hokke et al. 1996).

Biological Aspects

α1,3GT Gene

Mouse α1,3GT was first encoded by a multi-exon, single-copy gene which spans at least 80 kb. Exons 1–3 encode the 5′ untranslated sequence, whereas the protein coding sequence is distributed over exons 4–9. The start codon is located in exon 4, whereas exons 5–7 encode the “stem region.” The largest exon, exon 9, contains nearly all of the entire catalytic domain and in addition encodes 1.9 kb of the 3′ untranslated sequence. The mouse α1,3GT mRNA is alternatively spliced in the region that encodes the stem region (Joziasse et al. 1992). The stem region may play a role in determining the half-life of the enzyme within the cell (Cho et al. 1997). The porcine gene was also found to be organized in a similar manner (Sandrin et al.1994; Strahan et al. 1995). Sequencing of the clones demonstrated a single open reading frame coding for the predicted 371 amino acid protein sequence with a high homology to murine (75 % identity) and bovine (82 % identity) α1,3GT. Southern blot analyses showed the pig α1,3GT gene to be a single-copy gene, and northern analysis demonstrated an mRNA of 3.9 kb. By using fluorescence and isotopic in situ hybridization, the GGTA1 gene was mapped to the region q2.10–q2.11 of the pig chromosome 1.

α-Gal Epitope and Antibody

The anti-Gal antibody is present as a natural antibody and constitutes as much as 1 % of the circulating IgG in humans. A distinct evolutionary pattern has been reported, in which anti-Gal was found to be present in Old World monkeys and apes with titers comparable to those in humans, its corresponding antigenic epitope is expressed at relatively high levels by erythrocytes of New World monkeys. In further studies, an abundance of this α-Gal epitope (1 × 106−30 × 106) was found on cells from many species, including the kangaroo, mouse, rat, rabbit, pig, cow, horse, cat, dog, and dolphin (Galili et al. 1984, 1985), and on cells of prosimians, and of New World monkeys. In contrast, the catarrhines, which include Old World monkeys, apes, and humans, lack this enzyme activity because the α1,3GT gene is inactivated and in contrast produce large amounts of antibodies, designated as anti-Gal, against the α-Gal epitope, which is probably produced by constant antigenic stimulation by α-Gal-like epitopes which are located on the surface of normally occurring bacterial flora (Galili et al. 1987, 1988).

In terms of α1,3GT activity in individual organs, the mouse expresses a relatively lower α1,3GT activity than the pig in many organs, but the differences are in the tenfold range, except for the lung. The expression of α-Gal in adult pig islet cells is negligible. However, neonatal pig islets clearly express α-Gal (Rayat et al. 2003).

Knockout Mouse and Transgenic Mice

Transgenic mice with α1,3GT under the control of beta-actin promoter and cytomegalovirus enhancer have been produced. Compared with wild-type mice, the transgenic mice expressed GSI-B4 binding sites more intensely in the renal tubular brush border and lung alveolar epithelium and newly expressed them in the photoreceptor outer segments, goblet cells of the small intestine, and around spermatogonia. GSI-B4 binding sites were also detected in the liver of some transgenic mice. The transgenic mice tended to develop certain abnormalities, including more proteins in the urine, early death, a partial defect in hair growth, and low body weight (Ikematsu et al 1993).

Mouse α1,3GT that has been knocked out (KO) by homologous recombination appeared to develop normally, but develop eye cataracts within 4–6 weeks of birth (Thall et al. 1995; Tearle et al. 1996). Mating mice heterozygous for the inactivated α1,3GT resulted in a disturbance of the expected 1:2:1 ratio of wild type, heterozygote, and homozygote with a reduction in the transmission of the targeted allele.

Knockout Pig

On the other hand, regarding knocking out pig α1,3GT, gene targeting by homologous recombination in embryonic stem cells has not been feasible still now, since pig embryonic stem cells are not available for study. However, nuclear transplantation techniques from somatic cells to pig oocytes have been established at several institutes (Wilmut et al. 1997; Onishi et al. 2000). Piglets, in which one allele of the α1,3GT locus has been knocked out, were produced by nuclear transfer, using pig fibroblasts, by several different groups including our group. α1,3GT-KO pigs appear to be normal (Lai et al. 2002; Dai et al. 2002; Ramsoondar et al. 2003; Takahagi et al. 2005; Nottle et al. 2007). Serum anti-Gal IgM and IgG antibody levels were measured by ELISA in α1,3GT-KO pigs (78 estimations in 47 pigs). A low level of anti-Gal IgM was present soon after birth and rose to a peak at 4–6 m, which was maintained thereafter even in the oldest pigs tested (at >2 year). Anti-Gal IgG was also present at birth, peaked at 3 m, and after 6 m steadily decreased until almost undetectable at 20 m. No differences in this pattern were seen between pigs of different genders. Total IgM followed a similar pattern as anti-Gal IgM, but total IgG did not decrease after 6 m (Fang et al. 2012).

The studies of glycolipid antigens in GalT-KO pig tissues found increased levels of the uncapped LacNAc precursor, fucosylated blood group H type 2, the P1 antigen (Gala4nLc4), and the X2 antigen (by β1,3GalNAcT). However, these epitopes are not believed to mediate rejection because these are present on human cells. GalT-KO pigs did not produce new compensatory glycolipid compounds that react with human serum antibodies (Diswall et al. 2007, 2010). Lectin microarray analyses of adult pig islets from wild type and the GalT-KO pigs were carried out and the results compared to the corresponding values for human islets. In spite of the negligible expression of the Gal epitope on adult pig islets from wild type, α-linked GalNAc and Galβ1-3GalNAc were reduced in the adult islets from GalT-KO pigs. In a comparison between pigs and humans, the high-mannose form is originally rich in both APIs compared with humans, and human tissue appears to contain high levels of α-linked GalNAc (Miyagawa et al. 2013).

iGb3 Synthase

The iGb3 synthase (iGb3s: GT2) is encoded as a different gene from GT1 and has the potential for producing α-Gal epitopes. The gene can produce α-Gal antigens on a lactosyl core, thereby forming isoglobotriaosylceramide (iGb3; Galα3Galβ4Glc-NAcβ1ceramide) (Keusch et al. 2000). The rat expresses two distinct α1,3GT forms, GT1 and GT2, suggesting that the glycolipid iGb3 is produced by GT2 (Taylor et al. 2003). In addition, it was reported that GalT-KO mice still express Galα1,3Gal on lipid synthesized by GT2 (Milland et al. 2005). In addition, the transfection of iGb3s cDNA resulted in high levels of cell surface Galα1,3Gal synthesized via the isoglobo series pathway, thus demonstrating that mouse iGb3s is an additional enzyme capable of synthesizing the xenoreactive α-Gal epitope. α-Gal epitope synthesized by iGb3S, in contrast to α1,3GT, was resistant to downregulation by competition with α1,2FucT (Milland et al. 2006). On the other hand, using thin layer chromatography, the α-Gal epitope was not detected in tissues of GalT-KO pigs from their continuous reports. That is, on the basis of their findings, iGb3 was absent, and only fucosylated iGb3 was found (Diswall et al. 2007, 2010, 2011). Many studies related to the expression of α-Gal in GalT-KO pigs concluded that the available data do not support a relevant role for iGb3 in antibody-mediated pig-to-human xenotransplantation (Yung et al. 2009; Puga Yung et al. 2012). The issue of the existence of this GT2gene and its function in pigs has become less controversial.

Future Perspectives

α1,3GT-KO pigs with several human genes will become useful starting points for clinical xenotransplantation (Takahagi et al. 2005).

Since anti-Gal natural antibodies are produced by all humans, it can be exploited in a clinical setting to increase the immunogenicity of viral vaccines (Abdel-Motal et al. 2010) and autologous tumor vaccines that are processed to express α-Gal epitopes, by targeting such vaccines to antigen presenting cells (APC) at the vaccination site (Abdel-Motal et al. 2009).


Further Reading

  • Abdel-Motal et al. (2009)

  • Diswall et al. (2007)

  • Takahagi et al. (2005)

  • Zhang et al. (2004)


  1. Abdel-Motal UM, Wigglesworth K, Galili U (2009) Intratumoral injection of alpha-gal glycolipids induces a protective anti-tumor T cell response which overcomes Treg activity. Cancer Immunol Immunother 58:1545–1556PubMedCentralPubMedCrossRefGoogle Scholar
  2. Abdel-Motal UM, Wang S, Awad A, Lu S, Wigglesworth K, Galili U (2010) Increased immunogenicity of HIV-1 p24 and gp120 following immunization with gp120/p24 fusion protein vaccine expressing alpha-gal epitopes. Vaccine 28:1758–1765PubMedCentralPubMedCrossRefGoogle Scholar
  3. Blake DA, Goldstein IJ (1981) An alpha-d-galactosyltransferase activity in Ehrlich ascites tumor cells. Biosynthesis and characterization of a trisaccharide (alpha-d-galactose-(1 goes to 3)-N-acetyllactosamine). J Biol Chem 256:5387–5393PubMedGoogle Scholar
  4. Blanken WM, Van den Eijnden DH (1985) Biosynthesis of terminal Gal alpha 1–3Gal beta 1–4GlcNAc-R oligosaccharide sequences on glycoconjugates. Purification and acceptor specificity of a UDP-Gal:N-acetyllactosaminide alpha 1–3-galactosyltransferase from calf thymus. J Biol Chem 260:12927–12934PubMedGoogle Scholar
  5. Cho SK, Yeh JC, Cummings RD (1997) Secretion of alpha1,3-galactosyltransferase by cultured cells and presence of enzyme in animal sera. Glycoconj J 14:809–819PubMedCrossRefGoogle Scholar
  6. Dai Y, Vaught TD, Boone J, Chen SH, Phelps CJ, Ball S, Monahan JA, Jobst PM, McCreath KJ, Lamborn AE, Cowell-Lucero JL, Wells KD, Colman A, Polejaeva IA, Ayares DL (2002) Targeted disruption of the alpha1,3-galactosyltransferase gene in cloned pigs. Nat Biotechnol 20:251–255PubMedCrossRefGoogle Scholar
  7. Diswall M, Angström J, Schuurman HJ, Dor FJ, Rydberg L, Breimer ME (2007) Studies on glycolipid antigens in small intestine and pancreas from alpha1, 3-galactosyltransferase knockout miniature swine. Transplantation 84:1348–1356PubMedCrossRefGoogle Scholar
  8. Diswall M, Angström J, Karlsson H, Phelps CJ, Ayares D, Teneberg S, Breimer ME (2010) Structural characterization of alpha1,3-galactosyltransferase knockout pig heart and kidney glycolipids and their reactivity with human and baboon antibodies. Xenotransplantation 17:48–60PubMedCrossRefGoogle Scholar
  9. Diswall M, Gustafsson A, Holgersson J, Sandrin MS, Breimer ME (2011) Antigen-binding specificity of anti-αGal reagents determined by solid-phase glycolipid-binding assays. A complete lack of αGal glycolipid reactivity in α1,3GalT-KO pig small intestine. Xenotransplantation 18:28–39PubMedCrossRefGoogle Scholar
  10. Egge H, Kordowicz M, Peter-Katalinic J, Hanfland P (1985) Immunochemistry of I/i-active oligo- and polyglycosylceramides from rabbit erythrocyte membranes. Characterization of linear, di-, and triantennary neolactoglycosphingolipids. J Biol Chem 260:4927–4935PubMedGoogle Scholar
  11. Eto T, Ichikawa Y, Nishimura K, Ando S, Yamakawa T (1968) Chemistry of lipid of the posthemyolytic residue or stroma of erythrocytes. XVI. Occurrence of ceramide pentasaccharide in the membrane of erythrocytes and reticulocytes of rabbit. J Biochem (Tokyo) 64:205–213Google Scholar
  12. Fang J, Walters A, Hara H, Long C, Yeh P, Ayares D, Cooper DK, Bianchi J (2012) Anti-gal antibodies in α1,3-galactosyltransferase gene-knockout pigs. Xenotransplantation 19:305–310PubMedCentralPubMedCrossRefGoogle Scholar
  13. Galili U, Rachmilewitz EA, Peleg A, Flechner I (1984) A unique natural human IgG antibody with anti-alpha-galactosyl specificity. J Exp Med 160:1519–1531PubMedCrossRefGoogle Scholar
  14. Galili U, Macher BA, Buehler J, Shohet SB (1985) Human natural anti-alpha-galactosyl IgG. II. The specific recognition of alpha (1–3)-linked galactose residues. J Exp Med 162:573–582PubMedCrossRefGoogle Scholar
  15. Galili U, Clark MR, Shohet SB, Buehler J, Macher BA (1987) Evolutionary relationship between the natural anti-Gal antibody and the Gal alpha 1–3Gal epitope in primates. Proc Natl Acad Sci U S A 84:1369–1373PubMedCentralPubMedCrossRefGoogle Scholar
  16. Galili U, Shohet SB, Kobrin E, Stults CL, Macher BA (1988) Man, apes, and Old World monkeys differ from other mammals in the expression of alpha-galactosyl epitopes on nucleated cells. J Biol Chem 263:17755–17762PubMedGoogle Scholar
  17. Galili U, Swanson K (1991) Gene sequences suggest inactivation of alpha-1,3-galactosyltransferase in catarrhines after the divergence of apes from monkeys. Proc Natl Acad Sci USA 88:7401–7404PubMedCentralPubMedCrossRefGoogle Scholar
  18. Galili U, Tanemura M (1999) Significance of a-Gal (Gala1-3Galb1-4GlcNAc) Epitopes and a1,3Galactosyltranaferase in Xenotransplantation. Trends Glycosci Glycotechnol 11:317–327CrossRefGoogle Scholar
  19. Gastinel LN, Bignon C, Misra AK, Hindsgaul O, Shaper JH, Joziasse DH (2001) Bovine alpha1,3-galactosyltransferase catalytic domain structure and its relationship with ABO histo-blood group and glycosphingolipid glycosyltransferases. EMBO J 20:638–649PubMedCrossRefGoogle Scholar
  20. Henion TR, Macher BA, Anaraki F, Galili U (1994) Defining the minimal size of catalytically active primate alpha 1,3 galactosyltransferase: structure-function studies on the recombinant truncated enzyme. Glycobiology 4:193–201PubMedCrossRefGoogle Scholar
  21. Hokke CH, Zervosen A, Elling L, Joziasse DH, van den Eijnden DH (1996) One-pot enzymatic synthesis of the Gal alpha 1–>3Gal beta 1–>4GlcNAc sequence with in situ UDP-Gal regeneration. Glycoconj J 13:687–692PubMedCrossRefGoogle Scholar
  22. Honma K, Manabe H, Tomita M, Hamada A (1981) Isolation and partial structural characterization of macroglycolipid from rabbit erythrocyte membranes. J Biochem (Tokyo) 90:1187–1196Google Scholar
  23. Ikematsu S, Kaname T, Ozawa M, Yonezawa S, Sato E, Uehara F, Obama H, Yamamura K, Muramatsu T (1993) Transgenic mouse lines with ectopic expression of alpha-1,3-galactosyltransferase: production and characteristics. Glycobiology 3:575–580PubMedCrossRefGoogle Scholar
  24. Joziasse DH, Shaper JH, Van den Eijnden DH, Van Tunen AJ, Shaper NL (1989) Bovine alpha 1–3-galactosyltransferase: isolation and characterization of a cDNA clone. Identification of homologous sequences in human genomic DNA. J Biol Chem 264:14290–14297PubMedGoogle Scholar
  25. Joziasse DH, Shaper NL, Salyer LS, Van den Eijnden DH, van der Spoel AC, Shaper JH (1990) Alpha 1–3-galactosyltransferase: the use of recombinant enzyme for the synthesis of alpha-galactosylated glycoconjugates. Eur J Biochem 191:75–83PubMedCrossRefGoogle Scholar
  26. Joziasse DH, Shaper NL, Kim D, Van den Eijnden DH, Shaper JH (1992) Murine alpha 1,3-galactosyltransferase. A single gene locus specifies four isoforms of the enzyme by alternative splicing. J Biol Chem 267:5534–5541PubMedGoogle Scholar
  27. Keusch JJ, Manzella SM, Nyame KA, Cummings RD, Baenziger JU (2000) Expression cloning of a new member of the ABO blood group glycosyltransferases, iGb3 synthase, that directs the synthesis of isoglobo-glycosphingolipids. J Biol Chem 275:25308–25314PubMedCrossRefGoogle Scholar
  28. Lai L, Kolber-Simonds D, Park KW, Cheong HT, Greenstein JL, Im GS, Samuel M, Bonk A, Rieke A, Day BN, Murphy CN, Carter DB, Hawley RJ, Prather RS (2002) Production of alpha-1,3-galactosyltransferase knockout pigs by nuclear transfer cloning. Science 295:1089–1092PubMedCrossRefGoogle Scholar
  29. Larsen RD, Rajan VP, Ruff MM, Kukowska-Latallo J, Cummings RD, Lowe JB (1989) Isolation of a cDNA encoding a murine UDPgalactose:beta-d-galactosyl- 1,4-N-acetyl-d-glucosaminide alpha-1,3-galactosyltransferase: expression cloning by gene transfer. Proc Natl Acad Sci U S A 86:8227–8231PubMedCentralPubMedCrossRefGoogle Scholar
  30. Larsen RD, Rivera-Marrero CA, Ernst LK, Cummings RD, Lowe JB (1990) Frameshift and nonsense mutations in a human genomic sequence homologous to a murine UDP-Gal:beta-d-Gal(1,4)-d-GlcNAc alpha(1,3)-galactosyltransferase cDNA. J Biol Chem 265:7055–7061PubMedGoogle Scholar
  31. Lazarus BD, Milland J, Ramsland PA, Mouhtouris E, Sandrin MS (2002) Histidine 271 has a functional role in pig alpha-1,3galactosyltransferase enzyme activity. Glycobiology 12:793–802PubMedCrossRefGoogle Scholar
  32. Milland J, Christiansen D, Sandrin MS (2005) Alpha1,3-galactosyltransferase knockout pigs are available for xenotransplantation: are glycosyltransferases still relevant? Immunol Cell Biol 83:687–693PubMedCrossRefGoogle Scholar
  33. Milland J, Christiansen D, Lazarus BD, Taylor SG, Xing PX, Sandrin MS (2006) The molecular basis for galalpha(1,3)gal expression in animals with a deletion of the alpha1,3galactosyltransferase gene. J Immunol 176:2448–2454PubMedGoogle Scholar
  34. Miyagawa S, Maeda A, Takeishi S, Ueno T, Usui N, Matsumoto S, Teru Okitsu T, Goto M, Nagashima H (2013) A Lectin array analysis for wild-type and alpha-Gal-knockout pig islets, compared with humans. Surg TodayGoogle Scholar
  35. Molina P, Knegtel RM, Macher BA (2007) Site-directed mutagenesis of glutamate 317 of bovine alpha-1,3Galactosyltransferase and its effect on enzyme activity: implications for reaction mechanism. Biochim Biophys Acta 1770:1266–1273PubMedCentralPubMedCrossRefGoogle Scholar
  36. Nottle MB, Beebe LF, Harrison SJ, McIlfatrick SM, Ashman RJ, O’Connell PJ, Salvaris EJ, Fisicaro N, Pommey S, Cowan PJ, d’Apice AJ (2007) Production of homozygous alpha-1,3-galactosyltransferase knockout pigs by breeding and somatic cell nuclear transfer. Xenotransplantation 14:339–344PubMedCrossRefGoogle Scholar
  37. Onishi A, Iwamoto M, Akita T, Mikawa S, Takeda K, Awata T, Hanada H, Perry AC (2000) Pig cloning by microinjection of fetal fibroblast nuclei. Science 289:1188–1190PubMedCrossRefGoogle Scholar
  38. Puga Yung GL, Li Y, Borsig L, Millard AL, Karpova MB, Zhou D, Seebach JD (2012) Complete absence of the αGal xenoantigen and isoglobotrihexosylceramide in α1,3galactosyltransferase knock-out pigs. Xenotransplantation 19:196–206PubMedCentralPubMedCrossRefGoogle Scholar
  39. Ramsoondar JJ, Macháty Z, Costa C, Williams BL, Fodor WL, Bondioli KR (2003) Production of alpha 1,3-galactosyltransferase-knockout cloned pigs expressing human alpha 1,2-fucosylosyltransferase. Biol Reprod 69:437–445PubMedCrossRefGoogle Scholar
  40. Rayat GR, Rajotte RV, Hering BJ, Binette TM, Korbutt GS (2003) In vitro and in vivo expression of Galalpha-(1,3)Gal on porcine islet cells is age dependent. J Endocrinol 177:127–135PubMedCrossRefGoogle Scholar
  41. Sandrin MS, Dabkowski PL, Henning MM, Mouhtouris HE, McKenzie IFC (1994) Characterization of cDNA clones for porcine α(1,3)galactosyl transferase: the enzyme generating the Galα(1,3)Gal epitope. Xenotransplantation 1:81–88CrossRefGoogle Scholar
  42. Sepp A, Skacel P, Lindstedt R, Lechler RI (1997) Expression of alpha-1,3-galactose and other type 2 oligosaccharide structures in a porcine endothelial cell line transfected with human alpha-1,2-fucosyltransferase cDNA. J Biol Chem 272:23104–23110PubMedCrossRefGoogle Scholar
  43. Shah PS, Bizik F, Dukor RK, Qasba PK (2000) Active site studies of bovine alpha1–>3-galactosyltransferase and its secondary structure prediction. Biochim Biophys Acta 1480:222–234PubMedCrossRefGoogle Scholar
  44. Stellner K, Saito H, Hakomori S (1973) Determination of aminosugar linkages in glycolipids by methylation. Aminosugar linkages of ceramide pentasaccharides of rabbit erythrocytes and of Forssman antigen. Arch Biochem Biophys 155:464–472PubMedCrossRefGoogle Scholar
  45. Strahan KM, Gu F, Preece AF, Gustavsson I, Andersson L, Gustafsson K (1995) cDNA sequence and chromosome localization of pig alpha 1,3 galactosyltransferase. Immunogenetics 41:101–105PubMedCrossRefGoogle Scholar
  46. Sujino K, Malet C, Hindsgaul O, Palcic MM (1997) Acceptor hydroxyl group mapping for calf thymus alpha-(1–>3)-galactosyltransferase and enzymatic synthesis of alpha-d-Galp-(1–>3)-beta-d-Galp-(1–>4)-beta d-GlcpNAc analogs. Carbohydr Res 305:483–489PubMedCrossRefGoogle Scholar
  47. Takahagi Y, Fujimura T, Miyagawa S, Nagashima H, Shigehisa T, Shirakura R, Murakami H (2005) Production of alpha 1,3-galactosyltransferase gene knockout pigs expressing both human decay-accelerating factor and N-acetylglucosaminyltransferase III. Mol Reprod Dev 71:331–338PubMedCrossRefGoogle Scholar
  48. Taylor SG, McKenzie IF, Sandrin MS (2003) Characterization of the rat alpha(1,3)galactosyltransferase: evidence for two independent genes encoding glycosyltransferases that synthesize Galalpha(1,3)Gal by two separate glycosylation pathways. Glycobiology 13:327–337PubMedCrossRefGoogle Scholar
  49. Tearle RG, Tange MJ, Zannettino ZL, Katerelos M, Shinkel TA, Van Denderen BJ, Lonie AJ, Lyons I, Nottle MB, Cox T, Becker C, Peura AM, Wigley PL, Crawford RJ, Robins AJ, Pearse MJ, d’Apice AJ (1996) The alpha-1,3-galactosyltransferase knockout mouse. Implications for xenotransplantation. Transplantation 61:13–19PubMedCrossRefGoogle Scholar
  50. Thall AD, Malý P, Lowe JB (1995) Oocyte Gal alpha 1,3Gal epitopes implicated in sperm adhesion to the zona pellucida glycoprotein ZP3 are not required for fertilization in the mouse. J Biol Chem 270:21437–21440PubMedCrossRefGoogle Scholar
  51. Tumbale P, Jamaluddin H, Thiyagarajan N, Brew K, Acharya KR (2008) Structural basis of UDP-galactose binding by alpha-1,3-galactosyltransferase (alpha3GT): role of negative charge on aspartic acid 316 in structure and activity. Biochemistry 47:8711–8718PubMedCrossRefGoogle Scholar
  52. Van den Eijnden DH (2000) On the origin of oligosaccharide species. Glycosyltransferases in action. In: Ernst B, Hart G, Sinay P (eds) Oligosaccharides in chemistry and biology Part I, vol 2. Wiley/VCH, Weinheim, pp 589–624Google Scholar
  53. Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KH (1997) Viable offspring derived from fetal and adult mammalian cells. Nature 385:810–813PubMedCrossRefGoogle Scholar
  54. Yung GP, Schneider MK, Seebach JD (2009) Immune responses to alpha1,3galactosyltransferase knockout pigs. Curr Opin Organ Transplant 14:154–160CrossRefGoogle Scholar
  55. Zhang Y, Wang PG, Brew K (2001) Specificity and mechanism of metal ion activation in UDP-galactose:beta -galactoside-alpha-1,3-galactosyltransferase. J Biol Chem 276:11567–11574PubMedCrossRefGoogle Scholar
  56. Zhang Y, Swaminathan GJ, Deshpande A, Boix E, Natesh R, Xie Z, Acharya KR, Brew K (2003) Roles of individual enzyme-substrate interactions by alpha-1,3-galactosyltransferase in catalysis and specificity. Biochemistry 42:13512–13521PubMedCrossRefGoogle Scholar
  57. Zhang Y, Deshpande A, Xie Z, Natesh R, Acharya KR, Brew K (2004) Roles of active site tryptophans in substrate binding and catalysis by alpha-1,3 galactosyltransferase. Glycobiology 14:1295–1302PubMedCrossRefGoogle Scholar

Copyright information

© Springer Japan 2014

Authors and Affiliations

  1. 1.Division of Organ Transplantation, Department of SurgeryOsaka University Graduate School of MedicineOsakaJapan

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