Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.4.99.6 (sialyltransferase)
1,546 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Four common sialic acids (Sia), NeuAc, N-glycolyl-neuraminic acid (NeuGc), 4-O-acetyl-N-acetylneuraminic acid (4-O-Ac-NeuAc), and 9-O-Ac-NeuAc were examined for activation to their corresponding CMP-sialic acid conjugates and subsequently for their transfer to glycoprotein oligosaccharides by purified mammalian sialyltransferases. CMP-sialic acid synthetases from calf brain and from bovine and equine submaxillary glands were found to convert NeuAc, NeuGc, and 9-O-Ac-NeuAc to their corresponding CMP-sailic acids. In contrast, no conversion of 4-O-Ac-NeuAc to CMP-4-O-Ac-NeuAc was observed for any of the three synthetases examined. A new procedure for the preparation of CMP-9-O-Ac-NeuAc, CMP-NeuGc, and CMP-NeuAc in high yield and purity was developed, using the calf brain CMP-sialic acid synthetase. Each of these derivatives was tested as donor substrates for six mammalian sialyltransferases purified from porcine, rat, and bovine tissues, including a bovine GalNAc alpha 2,6 sialyltransferase whose purification is described in this report. The sialyltransferases examined represent those which form the Sia alpha 2,6Gal beta 1,4-GlcNAc-, Sia alpha 2,3Gal beta 1,3(4)GlcNAc-, Sia alpha 2,3Gal beta 1,3-GalNAc- and Sia alpha 2,6GalNAc- sequences found on N-linked and O-linked oligosaccharides of glycoproteins. CMP-NeuAc and CMP-NeuGc were equally good donor substrates for all six sialyltransferases. However, transfer of 9-O-Ac-NeuAc from CMP-9-O-Ac-NeuAc varied from only 10% to nearly 70% that of the transfer of NeuAc from CMP-NeuAc. Results are viewed to define the relative roles of direct transfer of these sialic acids and modification of glycosidically bound NeuAc in glycoproteins.
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PMID:Sialylation of glycoprotein oligosaccharides with N-acetyl-, N-glycolyl-, and N-O-diacetylneuraminic acids. 401 57

Fetal calf liver microsomes were found to be capable of sialylating 14C-galactosylated ovine submaxillary asialomucin. The main oligosaccharide product chain could be obtained by beta-elimination under reductive conditions and was identified as NeuAc alpha 2 leads to 3Gal beta 1 leads to 3GalNAcol (where GalNAcol represents N-acetylgalactosaminitol) by means of high performance liquid chromatography (HPLC) analysis and methylation. The branched trisaccharide Gal beta 1 leads to 3(NeuAc alpha 2 leads to 6)-GalNAcol and the disaccharide NeuAc alpha 2 leads to 6GalNAcol were not formed. Very similar results were obtained when asialofetuin and antifreeze glycoprotein were used as an acceptor. When 3H-sialylated antifreeze glycoprotein ([3H]NeuAc alpha 2 leads to 3Gal beta 1 leads to 3GalNAc-protein) was incubated with fetal calf liver microsomes and CMP-[14C]NeuAc, a reduced tetrasaccharide could be isolated. The structure of this product chain appeared to be [3H]NeuAc alpha 2 leads to 3Gal beta 1 leads to 3([14C]NeuAc alpha 2 leads to 6)GalNAcol, as established by means of HPLC analysis, specific enzymatic degradation with Newcastle disease virus neuraminidase, and periodate oxidation. These data indicate that fetal calf liver contains two sialyltransferases involved in the biosynthesis of the O-linked bisialotetrasaccharide chain. The first enzyme is a beta-galactoside alpha 2 leads to 3 sialyltransferase which converts Gal beta 1 leads to 3 GalNAc chains to the substrate for the second enzyme, a (NeuAc alpha 2 leads to 3Gal beta 1 leads to 3)GalNAc-protein alpha 2 leads to 6 sialyltransferase. The latter enzyme does not sialylate GalNAc or Gal beta 1 leads to 3GalNAc units but is capable of transferring sialic acid to C-6 of GalNAc in NeuAc alpha 2 leads to 3Gal beta 1 leads to 3GalNAc trisaccharide side chains, thereby dictating a strictly ordered sequence of sialylation of the Gal beta 1 leads to 3 GalNAc units in fetal calf liver.
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PMID:Biosynthesis of the O-glycosidically linked oligosaccharide chains of fetuin. Indications for an alpha-N-acetylgalactosaminide alpha 2 leads to 6 sialyltransferase with a narrow acceptor specificity in fetal calf liver. 619 Aug 2

The specificity of fetal calf liver and ovine and porcine submaxillary gland N-acetylgalactosaminide alpha 2 leads to 6 sialyltransferase was investigated with acceptors of low and high molecular weight containing O-glycosidically linked carbohydrate chains. It appeared that fetal calf liver microsomes were able to transfer sialic acid to C-6 of GalNAc in NeuAc(alpha 2 leads to 3)Gal(beta 1 leads to 3)GalNAc-R, in which the aglycon could be protein as well as p-nitrophenol (Nph). The substrates Gal(beta 1 leads to 3)GalNAc-R and GalNAc-R were inactive as acceptor with this enzyme source. Ovine and porcine submaxillary gland microsomes were both active with GalNAc-protein, Gal(beta 1 leads to 3)GalNAc-protein, NeuAc[alpha 2 leads to 3)Gal(beta 1 leads to 3)GalNAc-protein and NeuAc(alpha 2 leads to 3)Gal(beta 1 leads to 3)GalNAc alpha-Nph, but not with GalNAc alpha-Nph and Gal(beta 1 leads to 3)GalNAc alpha-Nph. The N-acetylgalactosaminide alpha 2 leads to 6 sialyltransferase which had been purified to homogeneity from porcine submaxillary gland [Sadler, J. E., Rearick, J. I. and Hill, R. L. (1979) J. Biol. Chem. 254, 5934-5941], was able to sialylate all three protein acceptors, but was virtually inactive with each of the three p-nitrophenyl glycosides. Our studies indicate that two N-acetylgalactosaminide alpha 2 leads to 6 sialtransferases exist acting on O-glycosidically linked carbohydrate chains. The first enzyme, present in fetal calf liver, has a narrow specificity with regard to the oligosaccharide structure, but shows no specificity for the aglycon. Based on its specificity this enzyme can be designated as an [alpha-N-acetylneuraminosyl 2 leads to 3-beta-galactosyl 1 leads to 3]-alpha-N-acetylgalactosaminide alpha 2 leads to 6 sialyltransferase. The second enzyme, present in porcine submaxillary gland, has an absolute requirement for protein as the aglycon. Once this condition is fulfilled, the enzyme is able to transfer sialic acid to each of the three oligosaccharide chains and thus the enzyme is an alpha-N-acetylgalactosaminylprotein alpha 2 leads to 6 sialyltransferase. The data also seem to suggest that ovine and porcine submaxillary gland microsomes contain, in addition to the latter enzyme activity, the alpha 2 leads to 6 sialyltransferase with the strict oligosaccharide specificity.
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PMID:Aglycon specificity of fetal calf liver and ovine and porcine submaxillary gland alpha-N-acetylgalactosaminide alpha 2 leads to 6 sialyltransferase. 661 53

A method has been developed to determine the activities of specific sialyltransferases by analysis of the products of the reaction. This method, which utilizes high performance liquid chromatography, distinguishes addition of sialic acid to the N-acetylgalactosamine vs. galactose residues of the mucin disaccharide Gal beta(1 leads to 3)GalNac, and can be used to distinguish formation of the 3'- and 6'-isomers of sialyllactose. For the bovine, ovine, and porcine submaxillary extracts, more than 95% of the activity with asialo ovine submaxillary mucin is due to formation of NeuAc alpha(2 leads to 6)GalNAc. With lactose as the acceptor, more than 95% of the alpha(2 leads to 3) isomer is produced. Activity with asialofetuin is due solely to the O-linked chain, with relative activity toward the galactose vs. GalNAc residues of 0.32, 1.5, and 0.10 for bovine, ovine, and porcine, respectively. The rat submaxillary gland extract showed equal formation of 3'- and 6'-sialyllactose, and very low activity with asialo ovine submaxillary mucin. However, at least 40% of the activity toward the Gal beta(1 leads to 3)GalNAc disaccharide of asialofetuin was directed toward the GalNAc residue. The relative preference of the N-acetylgalactosaminide alpha(2 leads to 6) sialyltransferase for a monosaccharide vs. a substituted GalNAc may play a role in regulation of chain length during mucin synthesis.
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PMID:Specificity of submaxillary gland sialyltransferases. 663 63

A beta-D-galactoside alpha 2 leads to 6 sialyltransferase was purified 500-fold in 14% yield from 14-day embryonic chicken liver. Characterization of the product of the sialyltransferase catalysis was accomplished by separation and permethylation of double-labelled ([14C]NeuAc, [3H]Gal) oligosaccharides following their release from the glycoprotein fetuin by hydrazinolysis. The enzyme transfers NeuAc to Gal(beta 1 leads to 4)GlcNAc(beta 1 leads to)R-terminated oligosaccharides; no activity was found towards Gal(beta 1 leads to 3)GalNAc(alpha 1 leads to)R structures. The trisaccharide. NeuAc(alpha 2 leads to 6)Gal(beta 1 leads to 4)Glc, was shown to be a good inhibitor of the sialyltransferase. Kinetic investigations of the enzyme indicate it to have a sequential, random bi-bi mechanism.
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PMID:Kinetic parameters of a beta-D-galactoside alpha 2 leads to 6 sialyltransferase from embryonic chicken liver. 675 14

A human hepatoma cell line (SK-H-MA) released a large amount of sialyltransferase (ST) and galactosyltransferase (GT) into the culture medium, whereas cells derived from normal human liver (Chang) released a large amount of GT but very little ST. The characteristics of hepatoma GT were studied since an abnormal GT isoenzyme has been associated with human gastrointestinal neoplasms. Both hepatoma and Chang medium GT activities had an absolute requirement for MnCl2 (25 mmol/l) and a broad optimal pH between 6.5 and 7.0, and were not affected by 0.1% Triton X-100. These two enzyme preparations were inhibited to the same extent by N-acetylglucosamine and N-acetylgalactosamine, while N-acetylglucosamine was 100 times more potent than N-acetylgalactosamine. Various nucleotides inhibited both enzyme activities equally well. Uracil-containing nucleotides were better inhibitors than thymine-containing nucleotides, and other nucleotides were only slightly inhibitory. The most effective inhibitor was UDP. More of the GT activity in hepatoma medium (65%) as compared to Chang medium (35%) bound to concanavalin A-Sepharose, and was eluted with 2.5% alpha-methylmannoside. These results suggest that the GTs from hepatoma and Chang media are not different in their enzymatic activity but may differ in their carbohydrate contents, which may be another manifestation of the neoplastic nature of the hepatoma cell line.
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PMID:Characterization of galactosyltransferase released from human hepatoma cells. 681 23

Rat liver Golgi was found to contain a sialyltransferase activity which would convert lacto-N-tetraose (Gal beta 1 goes to 3GlcNAc beta 1 goes to 3Gal beta 1 goes to 4Glc) to LS-tetrasaccharide a (NeuAc alpha 2 goes to 3Gal beta 1 goes to 3GlcNAc beta 1 goes to 3Gal beta 1 goes to 4Glc). The enzyme has been partially purified by affinity chromatography on CDP-hexanolamine agarose. Of the glycoprotein substrates examined, it utilizes the Gal beta 1 goes to 3GlcNAc sequence found on the asparagine-linked oligosaccharides of prothrombin as its preferred acceptor substrate, and thus has been tentatively designated a Gal beta 1 goes to 3GlcNAc alpha 2 goes to 3 sialyltransferase. The partially purified enzyme has an acceptor specificity distinct from other purified mammalian sialyltransferases which synthesize the NeuAc alpha 2 goes to 3Gal beta 1 goes to 3 GalNAc and NeuAc alpha 2 goes to 6 GalNAc sequences common to oligosaccharides O-linked to threonine or serine and the NeuAc alpha 2 goes to 6Gal beta 1 goes to 4GlcNAc sequence found on oligosaccharides N-linked to asparagine.
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PMID:Identification of a Gal beta 1 goes to 3GlcNAc alpha 2 goes to 3 sialyltransferase in rat liver. 706 22

High-pressure liquid chromatography was used to identify the sialo-oligosaccharide products obtained after sialylation of [14C]Gal-GalNAc-protein in vitro by an ovine submaxillary-gland microsomal fraction. Among other products, two isomeric trisaccharides could be identified. NeuAc alpha 2 leads to 3Gal beta 1 leads to 3GalNAcol and Gal beta 1 leads to 3-(NeuAc alpha 2 leads to 6)GalNAcol respectively, indicating that ovine submaxillary gland contains two sialyltransferases acting on mucin-type acceptors, a beta-galactoside alpha 2 leads to 3 sialyltransferase and a N-acetylgalactosaminide alpha 2 leads to 6 sialyltransferase. This conclusion was fully supported by methylation analysis of the two trisaccharide products.
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PMID:Specificity of ovine submaxillary-gland sialyltransferases. Application of high-pressure liquid chromatography in the identification of sialo-oligosaccharide products. 708 99

A sialyltransferase activity present in 7- to 12-day-old embryonic chicken brain catalyzes the transfer of sialic acid from CMP-sialic acid to the terminal galactose residue of [3H]nLcOse4Cer ([3H]Gal(beta 1-4).GlcNAc(beta 1-3)Gal(beta 1-4)Glc-Cer) to form NeuAc(alpha 2-3)-[3H]nLcOse4Cer (LM1 ganglioside). The product is sialidase-labile (96%), and the NeuAc group is linked to O-3 of the terminal galactose residue. The (alpha 2-3) linkage between sialic acid and the terminal galactose was determined on the basis of identification of 2,4,6-tri-O-methyl[3H]galactose obtained after hydrolysis of the permethylated enzymatic product. The CMP-sialic acid:nLcOse4Cer (alpha 2-3)sialyltransferase activity sediments (90%) at the junction of 1.2 M and 1.5 M on a discontinuous sucrose density gradient when still membrane bound (insoluble in 0.2% Triton X-100). The enzyme preparation also catalyzes the transfer of sialic acid from CMP-sialic acid to O-3 of GgOse4Cer (Gal(beta 1-3)GalNAc(beta 1-4)Gal(beta 1-4)Glc-Cer) to form NeuAc (alpha 2-3)GgOse4Cer (GM1b). Substrate inhibition studies indicate that these two reactions are probably catalyzed by the same enzyme.
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PMID:Biosynthesis in vitro of sialyl(alpha 2-3)neolactotetraosylceramide by a sialyltransferase from embryonic chicken brain. 713 Jan 78

Ovine submaxillary asialo-mucin was [14C]sialylated in vitro using a porcine liver cell-free preparation. The oligosaccharide chains were cleaved from the product glycoprotein by beta-elimination under reductive conditions, fractionated by gel filtration on Bio-Gel P-2 and characterized by thin-layer chromatography. The structure of the product chain was studied by periodate oxidation and analysis of the peeling products formed in the beta-elimination step. It appeared that [14C]-sialic acid had been introduced exclusively to the galactose residues of Gal beta(1 leads to 3)GalNAc disaccharide units occurring on the mucin as minor chains. No indication for a transfer to GalNAc residues on this glycoprotein was obtained. In agreement with this result sialyltransferase activities of porcine, rat, human and canine liver with Gal beta (1 leads to 3)GalNAc-protein acceptors were invariably much higher than those with ovine submaxillary asialo-mucin. When the asialo-mucin had been [14C]sialylated by an ovine submaxillary gland cell-free preparation analysis of the product oligosaccharide chain revealed the introduction of [14C]sialic acid to position C-6 on the GalNAc residues. The specificity of this transfer was reflected by the very high sialyltransferase activities of gland preparations with Gal beta (1 leads to 3)GalNAc-protein as well as GalNAc-protein acceptors. Mixed enzyme experiments indicated that the difference in liver and gland ovine submaxillary asialo-mucin sialyltransferase activities was not due to the presence of a specific inhibitor in the liver or an activator in the gland. It is concluded that porcine liver and likely liver of rat, man and dog contains a Gal beta (1 leads to 3)GalNAc-protein sialyltransferase, which is involved in the sialylation of O-glycosidically linked carbohydrate chains on serum glycoproteins. GalNAc-protein sialyltransferase activity, which richly occurs in ovine submaxillary gland, however, appears to be lacking from liver tissue.
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PMID:Specificity of sialyltransferase: sialylation of ovine submaxillary mucin in vitro. 721 66


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