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

---

Query: EC:3.1.1.53 (sialidase)
2,694 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Human liver sialidase was measured using methylumbelliferyl-N-acetylneuraminic acid as a substrate. The enzyme activity was linear for only 20 min and linearity was not improved by adding albumin, CaCl2, dithiothreitol, or Ep-459. The optimal pH was 4.5 and the apparent Km value, approximately 0.090 mM. Without substrate addition, the enzyme was unstable at temperatures between 0 and 37 degrees C, retaining only 35 and 5% of its activity, respectively, after 81/2 hr, but was protected by albumin at 5 mg/ml. The enzyme was more stable when either total liver or liver homogenate was kept frozen at -20 degrees C. Liver sialidase also retained about 70% of its activity after mechanical homogenization for 5 min. Potential inhibitors, notably, p- aminooxanilic acid, fetuin III, Triton X-100, mucin, sialyllactose, colominic acid, sodium taurocholate, N-acetylneuraminic acid, and methoxyphenyl-N-acetylneuraminic acid, were tested. Sialyllactose, methoxyphenyl-N-acetylneuraminic acid, fetuin, N-acetylneuraminic acid, and colominic acid were competitive inhibitors with Ki values of 1.12, 0.37, 0.20, 0.78, and 0.22 mM, respectively. The 0.11 M solutions of NaCl, LiCl, and KCl inhibited 20-30%, and CaCl2 about 60%, of the enzyme activity.
...
PMID:Methylumbelliferyl-N-acetylneuraminic acid sialidase in human liver. 647 33

A mixture of N-acetyl-[4,5,6,7,8,9-14C]neuraminosyl-alpha (2-3(6]-galactosyl-beta (1-4-glucose[( 14C]sialyl-lactose) and N-acetylneuraminosyl-alpha (2-3(6]-galactosyl-beta(1-4)-glucit-1-[3H]ol(sialyl-[3H]lactitol) as well as porcine submandibular gland mucin labeled with N-acetyl- and N-glycoloyl-[9-(3)H]neuraminic acid were administered orally to mice. The distribution of the different isotopes was followed in blood, tissues and excretion products of the animals. One half of the [14C]sialyl-lactose/sialyl-[3H]lactitol mixture given orally was excreted unchanged in the urine. The other half was hydrolysed by sialidase and partly metabolized further, followed by the excretion of 30% of the 14C-radioactivity as free N-acetyl-[4,5,6,7,8,9-14C]neuraminic acid and 60% of this radioactivity in the form of non-anionic compounds including expired 14CO2 within 24 h. The 14C-radioactivity derived from the [14C]sialyl-lactose/sialyl-[3H]lactitol mixture which remained in the bodies of fasted mice after 24 h was less than 1%. In the case of well-fed mice, a higher amount of the sialic acid residues was metabolized. The bulk of radioactivity of the mucin was resorbed within 24 h. About 40% of the radioactivity administered was excreted by the urine within 48 h; 30% of this radioactivity represented sialic acid and 70% other anionic and non-anionic metabolic products. 60% of the radioactivity administered remained in the body, and bound 3H-labeled sialic acids were isolated from liver. Sialyl-alpha (2-3)-[3H]lactitol was injected intravenously into rats; the substance was rapidly excreted in the urine without decomposition. These studies show that part of the sialic acids bound to oligosaccharides and glycoproteins can be hydrolysed in intestine by sialidase and be resorbed. This is followed either by excretion as free sialic acid or by metabolization at variable degrees, which apparently depends on the compound fed and on the retention time in the digestive tract.
...
PMID:Metabolism of sialic acids from exogenously administered sialyllactose and mucin in mouse and rat. 652 81

gpL115 is a lymphocyte surface component that is deficient in patients with the X-chromosome-linked immune deficiency Wiskott-Aldrich syndrome (6). The glycoprotein nature of gpL115 is demonstrated through labeling in carbohydrate moieties by [3H]NaBH4 and its synthesis by lymphocytes through labeling with [35S]methionine. Native gpL115 adheres to wheat germ lectin-Sepharose and sialidase-treated gpL115 does not adhere, indicating that native gpL115 adheres via clusters of sialic acid residues. When tested on peanut lectin, which shows specificity for the disaccharide Gal beta 1-3GalNAc, gpL115 is nonadherent and sialidase-treated gpL115 is adherent, indicating the presence of the sequence sialic acid-Gal beta 1-3GalNAc, which is characteristic for O-linked (mucin-type, acidic-type) carbohydrates. A surface glycoprotein with all the above characteristics was found on the lymphoblastoid cell line CEM. CEM cells were used as immunogen to generate the monoclonal antibody L10, an IgG1, which binds native and sialidase-treated gpL115 . Sialidase-treatment of gpL115 significantly alters its physical properties, reducing its electrophoretic mobility and changing its behavior on isoelectrofocusing. Cumulatively, these findings indicate that gpL115 , like glycophorin of erythrocytes and GPIb of platelets, is a sialoglyco protein with significant quantities of O-linked carbohydrate. On treatment with limiting sialidase concentrations, gpL115 of normal lymphocytes is transformed into a series of partially desialylated species of decreasing electrophoretic mobility. This finding resembles the situation with lymphocytes of some Wiskott-Aldrich syndrome patients. Lymphocytes of eight Wiskott-Aldrich syndrome patients were found to be deficient in 125I-labeled gpL115 . Lymphocytes from three of these patients displayed an abnormal 125I-component of apparent mol wt 135,000.
...
PMID:Characterization of a human lymphocyte surface sialoglycoprotein that is defective in Wiskott-Aldrich syndrome. 654 60

Purified liver lysosomes, prepared from rats previously injected with Triton WR-1339, exhibited sialidase activity towards sialyllactose, fetuin, submaxillary mucin (bovine) and gangliosides, and could be disrupted hypotonically with little loss in these activities. After centrifugation, the activities with sialyllactose and fetuin were largely recovered in the supernatant, demonstrating that they were originally in the intralysosomal space. The activities towards submaxillary mucin and gangliosides, on the other hand, remained in the pellet. In the supernatant, activity with fetuin or orosomucoid was markedly reduced by protease inhibitors, suggesting that proteolysis of these glycoproteins may be prerequisite to sialidase activity. The intralysosomal sialidase was solubilized from the mitochondrial-lysosomal fraction of rat liver and partially purified by Sephadex G-200, or Sephadex G-200 followed by CM-cellulose. The enzyme was maximally active at pH 4.7 with sialyllactose as substrate and had a minimum relative molecular mass of 60 000 +/- 5000 by gel filtration; it hydrolyzed a variety of sialooligosaccharides , those containing (alpha 2----3)sialyl linkages being better substrates than those with (alpha 2----6)sialyl linkages. The enzyme failed to attack submaxillary mucin and gangliosides. It was also inactive towards fetuin, orosomucoid and transferrin but capable of hydrolyzing glycopeptides from pronase digest of fetuin. In contrast to the intralysosomal sialidase, the sialidase partially purified from rat liver cytosol by (NH4)2SO4 fractionation followed by chromatography on DEAE-cellulose and CM-cellulose hydrolyzed fetuin and orosomucoid to the extent about half that for sialyllactose. The enzyme was maximally active at pH 5.8 and had a relative molecular mass of approximately 60 000. It also hydrolyzed gangliosides but not submaxillary mucin.
...
PMID:Rat-liver lysosomal sialidase. Solubilization, substrate specificity and comparison with the cytosolic sialidase. 672 66

A sialidase was solubilized with the aid of Triton X-100 from the insoluble material of a leucocyte homogenate. The enzyme was purified almost to homogeneity by chromatography on Sephadex G-75, equine submandibular gland mucin bound to Sepharose 4B and on Sephacryl S-200. The purification factor was 40 based on an increase of the specific enzyme activity from the Triton X-100 extract (pure enzyme: 40 mU/mg protein). Isolation of the active enzyme required the presence of a proteinase inhibitor. The sialidase is monomeric and has an average molecular mass of 48500 Da, a pH optimum of 4.6, hydrolyses preferably glycoprotein (fetuin) and sialyllactose, is activated by Ca2 and inhibited by N-acetyl-2,3-dehydro-2- deoxyneuraminic acid ( Neu5Ac2en ), Hg2 and N-(4-nitrophenyl) oxamic acid. The relatively stable enzyme shows only low activity with gangliosides and no activity with 4-O-acetylated sialic acid bound glycosidically.
...
PMID:Isolation and characterization of an oligosaccharide- and glycoprotein-specific sialidase from human leucocytes. 673 53

A sialidase, acting on gangliosides and mucus glycoproteins (pH optimum 4.0-4.5) was solubilized by 1% Triton X-100 and short ultrasonication from a crude mitochondrial-lysosomal fraction isolated from human liver. The enzyme was enriched over 1000-fold with the aid of affinity chromatography on equine submandibular gland mucin bound to Sepharose and 20mM Tris/HCl buffer, pH 7.5, as eluent. The sialidase exhibited a molecular mass of about 200 000 Da on Sephadex G-200. On analytical gel electrophoresis, an enzymically active protein band of about 70 000 Da was observed. A sialidase acting on sialyllactose remained in the membrane fraction and could not be solubilized.
...
PMID:Solubilization and affinity chromatography of a sialidase from human liver. 714 14

We isolated Gram-positive circular bacterium HB1 from intestinal microflora showing resistance to colonization by Clostridium difficile in mice (Su et. al., 1986a,b). We studied its enzymatic capacity to degrade mucin the first potential barrier to implantation of strains in the intestine. Its biochemical characteristics, terminal metabolites and the electrophoretic profiles of proteins and DNA-DNA homology indicated that it was a strain of Clostridium cocleatum. This strain displayed numerous glucosidase activities which were assumed to play a role in the degradation of mucin oligosaccharide chains in the digestive tract. These enzymes included alpha- and beta-galactosidases, beta-glucosidase, beta-N-acetylglucosaminidase, sialidase and alpha-N-acetylgalactosaminidase.
...
PMID:Identification of a Clostridium cocleatum strain involved in an anti-Clostridium difficile barrier effect and determination of its mucin-degrading enzymes. 750 16

The sequence in the assembly of the functional unit of selectin ligands containing sulfate, sialic acid, and fucose and also tumor-associated O-glycan structures was studied by examining the specificities of alpha 2,3-sialyltransferases (ST). The first enzyme, porcine liver ST, was 57, 37, and 79% active (Km: 0.105, 0.420, and 0.200 mM), respectively, toward 6-sulfo, 6-sialyl, or 6-O-methyl derivatives of the Gal beta 1,3GalNAc alpha- unit; C-3 or C-6 substitution on Gal abolished sialylation. An acrylamide copolymer (MW approximately 40,000) containing approximately 40 T-haptens and asialo Cowper's gland mucin (MW approximately 200,000) containing approximately 48 T-haptens was 5-fold more active as an acceptor as compared to Gal beta 1, 3GalNAc alpha-O-Al on a molecular weight basis. The second enzyme, a cloned alpha-2,3-ST specific for lactose-based structure, was 70, 102, and 108% active (Km: 0.500, 0.210, and 0.330 mM), respectively, toward 6-sialyl, 6-sulfo, or 6-O-methyl derivatives of the Gal beta 1,3GlcNAc beta- unit; C-3 and C-6 substitution on Gal abolished sialylation. Gal beta 1,4GlcNAc beta- and its 6-sulfo derivative were approximately 20% active; the Lewis a structure, Gal beta 1,3- (Fuc alpha 1,4)GlcNAc beta-, was not an acceptor. The acrylamide copolymers containing approximately 40 units of Gal beta 1,3GlcNAc beta-, Gal beta 1,3(6-sulfo)GlcNAc beta-, or fetuin triantennary asialo or bovine IgG diantennary glycopeptides were respectively 5.9-, 5.4-, 0.7-, and 0.1-fold as active. A transfer of 7-9 mol of NeuAc per mole of the above copolymers was catalyzed by this ST, the sialyl linkage being susceptible to alpha 2,3-specific sialidase. A partially purified Colo 205 Lewis type (alpha 1, 3/4) fucosyltransferase catalyzed the formation of 3'-sialyl-6-sulfo Lewis a from [9-3H]NeuAc alpha 2, 3Gal beta 1, 3(6-sulfo)GlcNAc beta-O-Allyl and copolymer containing [9-3H]NeuAc alpha 2, 3Gal beta 1, 3(6-sulfo)GlcNAc beta- units, using GDP[14C]Fuc as fucosyl donor. The third enzyme, HL-60 ST, was 103% active with Gal beta 3(6-sulfo)GalNAc alpha- but was only 8% active with 6-sialo compound; it showed 11.6-fold greater activity with the copolymer of T-hapten. Further, we observed the alpha 2,3 sialylation of Gal beta 1,4GlcNAc beta- but not Gal beta 1,3GlcNAc beta- by HL60-ST, consistent with the occurrence of 3'-sialyl LacNAc and 3'-sialyl Lewis x units in leukosialin of HL60.(ABSTRACT TRUNCATED AT 400 WORDS)
...
PMID:Selectin ligands and tumor-associated carbohydrate structures: specificities of alpha 2,3-sialyltransferases in the assembly of 3'-sialyl-6-sialyl/sulfo Lewis a and x, 3'-sialyl-6'-sulfo Lewis x, and 3'-sialyl-6-sialyl/sulfo blood group T-hapten. 753 77

Glycophorin A was digested with glycoprotease (Pasteurella haemolytica) and the digest was fractionated by a combination of high-pressure column chromatographies to produce the glycopeptides GPA-1 to GPA-6. Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. The glycopeptides thus obtained were treated with sialidase and beta-galactosidase. The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 67%), 81% (calcd. 86%), and 50% (calcd. 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. These results indicate that clusters I and II react with the antibody to the same extent. The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin.
...
PMID:Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128). 768 97

A sialic acid-binding lectin, AchatininH (ATNH), having unique specificity towards 9-O-acetylneuraminic acid, has been purified and characterized. The specificity of this lectin for O-acetylsialic acids was studied in detail, using various sialic acid derivatives and sialoglycoproteins. The potent inhibition of hemagglutination by bovine submaxillary mucin (BSM), which contains 9(7,8)-O-acetylsialic acid and by free 9-O-acetylneuraminic acid confirms the preferential affinity towards this sugar. Further support for the role of O-acetylsialic acid was obtained by sialidase treatment of BSM. O-Deacetylation of the sialic acid residue abolished its inhibitory potency. Moreover, when the trihydroxypropyl side chain of the sialic acid molecule was modified by periodate-borohydride treatment, the truncated C7-sialic acid was unable to bind ATNH. This result suggests that the glycerol side chain of Neu5Ac, especially the C-8 and/or C-9 portion is an important determinant for ATNH. The hemagglutination-inhibition results using several mono-, di-, and tri-saccharides containing terminal sialic acid and various sialoglycoproteins reveals that ATNH preferentially binds the alpha-(2-->6)-linked sialic acid. Furthermore, beta-D-GlcNAc-(1-->3)-[alpha-NeuGc-(2-->6)]-GalNAc-ol was found to be the best ligand for ATNH.
...
PMID:The specificity of the binding site of AchatininH, a sialic acid-binding lectin from Achatina fulica. 773 61


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>

---