The combining site of the Bauhinia purpurea alba lectin was studied by quantitative precipitin and precipitin inhibition assays. Of 45 blood group substances, glycoproteins, and polysaccharides tested, 35 precipitated over 75% of the lectin. Precursor blood group substances with I activity (Cyst OG 10% from 20% and Cyst OG 20% from 10%), desialized fetuin, and desialized ovine salivary glycoprotein, in which more than 75% of the carbohydrate side chains have dGalN Ac linked through α1 → to the OH group of Ser or Thr of a protein core, completely precipitated the lectin. The poorly reactive blood group substances after mild acid hydrolysis or Smith degradation, as well as sialic acid-containing glycoproteins after removal of sialic acid, had substantially increased activity so that more than 80% of the lectin was precipitated. Precipitability with various blood group substances and glycoproteins is ascribable to the terminal nonreducing dGalNAc, dGalβ1 → 3dGalNAc, dGalβ1 → 3 or 4dGlcNAc, and dGalβ1 → 3 or 4dGlcNAcβ1 → 3dGal determinants on the carbohydrate moiety. Of the monosaccharides tested for inhibition of precipitation, dGalNAc and its p-nitrophenyl and methyl α-glycosides were best. These compounds were four to five times better than the corresponding dGal compounds but methyl βDGalNAcp was only about 40% more active than methyl βdGalp. The α-anomers of p-nitrophenyl DGalNAcp and dGalp, were twice as active as the corresponding β-anomers. Methyl αDGalNAcp was four times as active as the β-anomer but the inhibitory power of the methyl α- and β-anomers of dGal were about equal. Among the oligosaccharides tested, dGalβ1 → 3dGalNAc and its tosyl derivatives were most active, the tosyl glycosides being about twice as active as dGalβ1 → 3dGalNAc, which was somewhat more active than dGalNAcα1 → 6dGal and dGalNAc, and 2.5 and 5 times as active as dGalNAcα1 → 3dGalβ1 → 3dGlcNAc and dGalNAcαl → 3dGa1, respectively (blood group A specific). These findings suggest that a subterminal dGalNAc β-linked and substituted on carbon 3 plays an important role in binding. Consistent with this inference are the findings that dGalβ1 → 3dGlcNAc and dGalβ1 → 6dGal were poorer inhibitors although dGalβ1 → 3dGlcNAc was two to three times as active as glycosides of dGal. Oligosaccharides with terminal nonreducing dGal and subterminal α-linked dGal were as active or less active than dGal. dGalβ1 → 3dGlcNAcβ1 → 3dGalβ1 → 4dGlc (lacto-N-tetraose) and dGalβ1 → 3dGlcNAcβ1 → 3dGal-β1-O-(CH2)8COOCH3 were equally active and 1.5 times as potent as dGalβ1 → 3dGlcNAc whereas dGalβ1 → 3dGlcNAcβ1 → 6dGal was only 40% as potent as dGalβ1 → 3dGlcNAc suggesting that a third sugar may be part of the determinant. Substitution of dGalβ1 → 3dGlcNAcβ1 → 3dGalβ1 → 4dGlc on the subterminal dGlcNAc by lFucα1 → 4 in lacto-N-fucopentaose II reduced activity fourfold; if the nonreducing dGal is substituted by lFucα1 → 3 as in lacto-N-fucopentaose I its activity is almost completely abolished. This suggests that a terminal nonreducing dGal as well as subterminal dGlcNAc are contributing to binding. The β → 3 linkage of the terminal dGal to the subterminal amino sugar is significant since dGalβ1 → 4dGlcNAc is a poorer inhibitor. Although the available data suggest that the combining site of the lectin Bauhinia purpurea alba may be most complementary to the structure dGalβ1 → 3dGalNAcβ1 → 3dGal, several other possibilities remain to be tested when suitable oligosaccharides become available. 相似文献
Sophora japonica lectin agglutinates human B erythrocytes strongly and A1 erythrocytes weakly. Bivalent metal ions such as Ca2+, Mn2+, or Mg2+ were shown to be essential for hemagglutinating and precipitating activities. At optimal concentrations of bivalent metal ions, hemagglutinating activity was highest between pH 8.5 and 9.0 and decreased sharply below pH 8.5, whereas precipitating capacity was maximal between pH 6.7 and 9.5. The combining site of the S. japonica lectin was explored by quantitative precipitin and precipitin inhibition assays. This lectin showed substantial differences in precipitation with several blood group B substances ascribable to heterogeneity resulting from incomplete biosynthesis of their carbohydrate side chains. The lectin precipitated moderately well with A1 substance and precursor blood group I fractions (OG). It precipitated weakly or not at all with A2, H, or Lea substances. In inhibition assays, glycosides of dGalNAc were about five to six times better than those of dGal; dGalNAc itself was about six times better than dGal. Nitrophenyl glycosides were all substantially better than the methyl glycosides, indicating a hydrophobic contribution to the site subterminal to the nonreducing moiety. Although nitrophenyl β-glycosides were much better than the corresponding α-glycosides, the methyl α-and βDGalNAcp were equal in activity as were methyl α- and βDGalp. Among the oligosaccharides tested, the β-linked N-tosyl-l-serine glycoside of dGalβ1 → 3dGalNAc was best and was as active as p-nitrophenyl βDGalNAcp, whereas dGalβ1 → 3dGalNAc α-N-tosyl serine and the nitrophenyl and phenyl α-glycosides of dGalβ1 → 3dGalNAc were much less active, suggesting that the hydrophobic moiety and/or a subterminal dGalNAc β-linked and substituted on carbon 3 play an important role in binding and that a β-linked glycoside of dGalβ1 → 3dGalNAc may be an essential requirement for binding. The results of inhibition studies with other oligosaccharides indicate that a subterminal dGlcNAc substituted on carbon 3 or 4 by dGalβ may contribute somewhat to binding and that whether the dGlcNAc is linked β1 → 3 or β1 → 6 to a third sugar does not contribute to or interfere with binding. The β1 → 3 linkage of the terminal dGal to the subterminal amino sugar is significant since dGalβ1 → 4dGlcNAc was one-half as active as the corresponding β1 → 3-linked compound and the subterminal sugar must be unsubstituted for optimal binding. N-Acetyllactosamine was 50% more active than lactose, indicating that the subterminal N-acetamido group was also contributing significantly to binding. A variety of other sugars, glycosides, and oligosaccharides showed little or not activity. From the oligosaccharides available, the combining size of this lectin would appear to be least as large a β-linked disaccharide and most complementary to dGalβ1 → 3dGalNAc β-linked to tosyl-l-serine the most active compound tested. 相似文献
Carbohydrate structures in the interior of a blood group A active substance (MSS) were exposed by one and by two Smith degradations. Reactivities of the original glycoprotein and its Smith degraded products with 13 different lectins and with anti-I Ma were studied by quantitative precipitin assay. MSS and its first Smith degraded product completely precipitated Ricinus communis hemagglutinin with five times less of the first Smith degraded glycoprotein being required for 50% precipitation. The second Smith degraded material precipitated only 90% of the lectin. MSS did not precipitate peanut lectin, whereas its first and second Smith degraded products completely precipitated the lectin. The first Smith degraded glycoprotein also reacted well with Wistaria floribunda, Maclura pomifera, Bauhinia purpurea alba, and Geodia lectins indicating that its carbohydrate moiety could contain dGalNAc, dGalβ1 → 3dGalNAc, dGalβ1 → 4dGlcNAc, dGalβ1 → 3dGlcNAcβ1 → 3dGal and/or dGalβ1 → 4dGlcNAcβ1 → 6dGal and/or dGalβ1 → 4dGlcNAcβ1 → 6dGalNAc determinants at nonreducing ends. The second Smith degraded material precipitated well with Ricinus communis hemagglutinin, Arachis hypogaea, Geodia cydonium, Maclura pomifera, and Helix pomatia lectins showing that dGalNAc, dGalβ1 → 3dGalNAc, dGalβ1 → 4dGlcNAc residues at terminal nonreducing ends could be involved. Monoclonal anti-I Ma (group 1) serum reacted strongly with the first Smith degraded product indicating large numbers of anti-I Ma determinants, dGalβ1 → 4dGlcNAcβ1 → d 6dGal and/or dGalβ1 → 4dGlcNAcβ1 → 6dGalNAc at nonreducing ends. The comparable activities of the native and Smith degraded products with wheat germ lectin indicate capacity to react with DGlcNAc residues at nonreducing ends and/or at positions in the interior of the chain. The totality of lectin reactivities indicates heterogeneity of the carbohydrate side chains. Oligosaccharides with 3H at their reducing ends released from the protein core of the first and second Smith degraded products were obtained by treatment with 0.05 m NaOH and 1 M NaB3H4 at 50 °C for 16 h (Carlson degradation). The liberated reduced oligosaccharides were fractionated by dialysis, followed by retardion, Bio-Gel P-2, P-4, and P-6 columns. They were further purified on charcoal-celite columns, and by preparative paper chromatography and high-pressure liquid chromatography. Their distribution by size was estimated by the yields on dialysis, Bio-Gel P-2, and Bio-Gel P-6 chromatography, and from the radioactivity of the reduced sugars. Of the oligosaccharide fractions from the first Smith degraded product, about 77% of the carbohydrate side chain residues contained from 1 to 6 sugars, 13% from 7 to perhaps 12 sugars, and 10% was nondialyzable (polysaccharides and glycopeptide fragments). Of the second Smith degraded product, approximately 82% of carbohydrate residues had from 1 to 6 sugars, 14% from 7 to perhaps 20 sugars and 4% was nondialyzable. The biological activity profile of the two Smith degraded products together with the size distributions of the oligosaccharides indicated that their carbohydrate side chains, comprised a heterogeneous population ranging in size from 1 to about 12 sugars. When most of these chains that are shorter than hexasaccharides are fully characterized it may be possible to reconstruct the overall structure of the carbohydrate moiety of the blood group substances and account for their biological activities. 相似文献
The lectin from Euonymus europeus seeds was purified by adsorption onto insoluble polyleucyl hog A + H blood group substance and subsequent elution with lactose. The isolated lectin formed three lines in immunoelectrophoresis against rabbit antisera to the crude seed extract and showed three components on electrophoresis in acrylamide gel at pH 9.4. In analytical isoelectric focusing the purified lectin had six closely spaced bands with pI from 4.3 to 4.7. It sedimented as two peaks: a big symmetrical peak with s20,w0 of 7.8 and another small, diffuse moving peak. The intrinsic viscosity was 0.057 dl/g and the Mr calculated from the sedimentation coefficients, intrinsic viscosity, and V? of 0.71 was about 166,000. In sodium dodecyl sulfate, it gives subunits of Mr 17,000 and 35,000; 20% of the 35,000 subunit resists reduction by dithiothreitol in 7 m guanidine-HCl. The Euonymus lectin is a glycoprotein containing 4.8% d-galactose, 2.9% d-glucose, and 2.8% N-acetyl-d-glucosamine. The purified lectin precipitated well with B and H blood group substances and with the P1 fraction of blood group B substance but not with A1 substances. It precipitated poorly with Lea and Leb and precursor I blood group substances. Inhibition of precipitation with milk and blood group oligosaccharides showed the lectin to be most specific for blood group B oligosaccharides having the structure: dGalα1 → 3[lFucα1 → 2]dGalβ1 → 3 or 4dGlcNAcβ→. It is also inhibited by blood group H oligosaccharides but to a lesser degree. For 50% inhibition of precipitation, 3.5, 850, and 290,000 nmol of B and H oligosaccharides and lactose, respectively are required. The B and H specificities are an intrinsic property of a single lectin site since absorption and elution from an H immunoadsorbent gave material with B as well as H specificity. Millipore-filtered crude extracts of Euonymus europeus preserved with 0.02% sodium azide are stable in the refrigerator for many months and can be used for quantitative precipitin and for quantitative inhibition assays, results being the same as with purified lectin. 相似文献
The specificity of the purified soybean agglutinin has been studied immunochemically by quantitative precipitin and quantitative precipitin inhibition assays. The lectin is precipitated by human A and Lea blood-group substance, by the products of the second, third, fourth, and fifth stages of periodate oxidation of a human H blood-group substance (JS), and by precursor blood-group substances, as well as by a pig-submaxillary mucin having blood-group A activity, by partially hydrolyzed blood-group B substances (Pl fraction), and by group C streptococcal polysaccharide. The activity is attributable to terminal α-linked 2-acetamido-2-deoxy-d-galactopyranosyl or to α- or β-d-galactopyranosyl residues. The lectin did not precipitate with human blood-group H substances, with the product of the first stage of periodate oxidation (JS), with streptococcal group A polysaccharide, or with pig-submaxillary mucin devoid of blood-group A activity, and is poorly precipitated by blood-group B substances. Inhibition of precipitation with various monosaccharides indicated that the lectin is strongly specific for 2-acetamido-2-deoxy-d-galactose and for its oligosaccharides, and to a lesser extent for d-galactose and its oligosaccharides; the α-glycosides of both sugars were slightly more reactive than the β-glycosides of 2-acetamido-2-deoxy-d-galactose, and both α- and β-glycosides were more active than the free monosaccharides. Aromatic α- and β-glycosides of 2-acetamido-2-deoxy-d-galactose and d-galactose were better inhibitors than the corresponding methyl or ethyl compounds. The blood-group A trisaccharide α-d-GalNAcp-(1→3)-β-d-Galp-(1→3)-d-GlcNAc was more active than the disaccharide lectins by the use of precipitation with polysaccharides, as well as inhibition reactions, is essential to the understanding of their reactivity with cell-surface receptors. 相似文献
β-d-Gal-(1 → 4)-β-d-GlcNAc-OC6H4NO2-p (p-nitrophenyl N-acetyl-β-lactosaminide) and β-d-Gal-(1 → 6)-β-d-GlcNAc-OC6H4NO2-p (p-nitrophenyl N-acetyl-β-isolactosaminide) were regioselectively synthesized from lactose and p-nitrophenyl 2-acetamido-2-deoxy-glucopyranoside, employing transglycosylation by the β-d-galactosidase from Bacillus circulans and by controlling the concentration of organic solvent in the reaction system. The (1 → 4)-linked disaccharide was formed exclusively when the concentration of organic solvent was high, whereas the (1 → 6)-linked isomer was produced with a low concentration. Further utilization of the transglycosylation by the enzyme led to the regioselective formation of β-d-Gal-(1 → 4)-d-GalNAc and β-d-Gal-(1 → 4)-β-d-GalNAc-OC6H4NO2-p. With the enzyme, β-d-galactosyl transfer occurred preferentially at the O-4 position of GlcNAc and GalNAc, regardless of the configuration of the hydroxyl group. 相似文献
β-d-Mannosidase (β-d-mannoside mannohydrolase EC 3.2.1.25) was purified 160-fold from crude gut-solution of Helix pomatia by three chromatographic steps and then gave a single protein band (mol. wt. 94,000) on SDS-gel electrophoresis, and three protein bands (of almost identical isoelectric points) on thin-layer iso-electric focusing. Each of these protein bands had enzyme activity. The specific activity of the purified enzyme on p-nitrophenyl β-d-mannopyranoside was 1694 nkat/mg at 40° and it was devoid of α-d-mannosidase, β-d-galactosidase, 2-acet-amido-2-deoxy-d-glucosidase, (1→4)-β-d-mannanase, and (1→4)-β-d-glucanase activities, almost devoid of α-d-galactosidase activity, and contaminated with <0.02% of β-d-glucosidase activity. The purified enzyme had the same Km for borohydride-reduced β-d-manno-oligosaccharides of d.p. 3–5 (12.5mm). The initial rate of hydrolysis of (1→4)-linked β-d-manno-oligosaccharides of d.p. 2–5 and of reduced β-d-manno-oligosaccharides of d.p. 3–5 was the same, and o-nitrophenyl, methylumbelliferyl, and naphthyl β-d-mannopyranosides were readily hydrolysed. β-d-Mannobiose was hydrolysed at a rate ~25 times that of 61-α-d-galactosyl-β-d-mannobiose and 63-α-d-galactosyl-β-d-mannotetraose, and at ~90 times the rate for β-d-mannobi-itol. 相似文献
The sugar specificity of Escherichia coli 346 and of the type-1 fimbriae isolated from this organism has been studied by quantitative inhibition of the agglutination of mannan-containing yeast cells. The best inhibitors of the agglutination by the bacteria were the oligosaccharides Manα1→6[Manα1→3]Manα1→6[Manα1→2Manα1→3]ManαOMe, Manα1→6[Manα1→3]Manα1→6[Manα1→3]ManαOMe and Manα1→3Manβ1→4GlcNAc, and the aromatic glycoside p-nitrophenyl α-d-mannoside, all of which were 20–30 times more inhibitory than methyl α-d-mannoside. The disaccharides Manα1→3Man, Manα1→2Man and Manα1→6Man, the tetrasaccharide Manα1→2Manα1→3Manβ1→4GlcNAc and the pentasaccharide Manα1→2Manα1→2Manα1→3Manβ1→4GlcNAc, were all poor inhibitors. A very good correlation was found between the relative inhibitory activity of the different sugars tested with intact bacteria and with the isolated fimbriae. Our findings show that the combining site of the E. coli lectin is an extended one, corresponding to the size of a trisaccharide, that it contains a hydrophobic region, and that it is in the form of a pocket on the surface of the lectin. The combining site fits best the structures found in short oli gomannosidic chains present in N-glycosidically linked glycoproteins. 相似文献
The lectin II from Ulex europaeus seeds was purified by adsorption on insoluble polyleucyl hog A + H blood group substance and elution with 35% ethylene glycol, and by chromatography on ?-aminocaproyl-fucosyl-amine-agarose. In immunodiffusion against rabbit antiserum to the crude extract, the isolated lectin formed one line which fused with one of the five formed by crude extract. The purified lectin showed two bands on acrylamide electrophoresis under alkaline or acid conditions but only one band of molecular weight 23,000 if the electrophoresis was in the presence of 0.1% sodium dodecyl sulfate at pH 8.8. The agglutinating and precipitating abilities are abolished by EDTA and can be restored by bivalent cations. The purified lectin precipitated to different extents with blood group A1, A2, B, HLeb, Lea, and I precursor substances and with acid- or Smith-degraded substances. Inhibition of precipitation indicated that the lectin site was unusual in that it interacted most strongly with the h-specific oligosaccharide and with 2′-fucosyllactose, followed by β1 → 4 linked oligomers of dGlcNAc. Molecular models showed that all these inhibitors have a similarity in three-dimensional structures that could account for their activities. 相似文献
Four different H-type 1 (LedH) blood-group-active glycosphingolipids (LedH-I–IV) have been isolated from the plasma of blood-group O Le(a?b?) secretors. The agglutination of O Le(a?b?) erythrocytes from secretors by 50 μl of 4 hemagglutinating units of caprine anti-LedH (anti-H-type 1) serum was inhibited by 0.02 μg of each of all four glycolipids. No Lea or Leb activities or reaction against Ulex europaeus lectin could be found. LedH-I, -II, -III, and -IV at 0.05, 0.01, 0.01, and 0.02 μg each are sufficient for incubation in order to convert 9 × 107 O Le(a?b?) erythrocytes from nonsecretors into H-type 1 (LedH)-positive cells. Structural analysis of the H-type 1 glycolipids was performed in comparison to that of Lea- and Leb-blood-group-active glycolipids from human plasma isolated previously: Gas chromatography of peracetylated alditols revealed sugar composition. Combined gas chromatography-mass spectrometry established the glycosidic linkages. Together with the results obtained by direct inlet mass spectrometry of permethylated glycosphingolipids and by 360-MHz 1H nuclear magnetic resonance spectroscopy (Egge, H., and Hanfland, P., 1981, Arch. Biochem. Biophys., 210, 396–404; Dabrowski, J., Hanfland, P., Egge, H., and Dabrowski, U., 1981, Arch. Biochem. Biophys., 210, 405–411) the complete structures of the oligosaccharide chains of the Lea-, Leb-, and H-type 1-active glycolipids were established: Galβ1 → 3GlcNAc(4 ← 1αFuc)β1 → 3Galβ1 → 4Glcβ1 → 1 Cer for the Lea antigens; Fucα1 → 2Galβ1 → 3GlcNAc(4 ← 1αFuc)β1 → 3Galβ1 → 4Glcβ1 → 1 Cer for the Leb antigens; and Fucα1 → 2Galβ1 → 3GlcNAcβ1 → 3Galβ1 → 4Glcβ1 → 1 Cer for the H-type 1 (LedH) glycolipids. The diverse antigens of the same blood-group specificity obviously differ from one another in their lipid residue. In addition, plasmatic neolactotetraosylceramide could be identified, differing from that of human erythrocytes by a slower migration behavior in thin-layer chromatography. 相似文献
Preparations of the β-galactoside-binding lectin of bovine heart have been shown to stimulate the sialylation of the oligosaccharide Ga1β1→4G1cNAc and asialo-α1-acid glycoprotein by bovine colostrum β-D-galactoside α2→6 sialyltransferase. Kinetic data revealed that in the presence of lectin the Km values for Ga1β1→4G1cNAc and CMP-NeuAc were reduced from 25.0 to 11.6 mM and from 0.42 to 0.19 mM respectively, but the Km for asialo-α1-acid glycoprotein and the Vmax values for all three substrates were little affected. Stimulation by the lectin was partially inhibited by Fucα1→2Ga1β1→4G1cNAc. This, together with the effects of certain plant lectins, suggests that the stimulation of sialytransferase may be mediated through the carbohydrate-binding properties of the lectin. 相似文献
Periodate oxidation of LPG-1 established that N-acetylneuraminic acid residues are linked preponderantly α-(2→3) to D-galactose residues. The resistance of 2-acetamido-2-deoxyD-galactose residues to periodate oxidation suggests that they are linked at either O-3 or O-4 to D-galactose residues. After treatment of LPG-I with alkaline sulfite, ≈80% of 2-acetamido-2-deoxygalactose was recovered as the sulfonic acid derivative. The Gal→GalNAc disaccharide released from sialic-acid-free LPG-I by digestion with endo-2-acetamido-2-deoxy-α-D-galactosidase (which suggests an α-D-GalNAc→-L-Ser or -L-Thr linkage) gave a high color-yield in the Morgan—Elson reaction, indicating that 2-acetamido-2-deoxy-D-galactose residues are linked at C-3 to D-galactose residues. The migration of the released Gal-GalNAc disaccharide was the same as that of a standard sample of O-β-D-galactosyl-(1→3)-2-acetamido-2-deoxy-D-galactose. Treatment of sialic acid-free LPG-I with Streptococcus pneumoniae β-D-galactosidase, which hydrolyzes only galactosides linked β-D-(1→4) gave no free D-galactose, whereas treatment of LPG-I with bovine testes β-D-galactosidase released > 90% of D-galactose. These results provide evidence for β-D-Galp-(1→3)-α-D-GalNAcp-(1→3)-L-Ser or -L-Thr and α-NeuAc-(2→3)-β-D-Galp-(1→3)-α-D- GalNAcp-(1→3)-L-Ser or -L-Thr structures. The sensitivity of the methods used and the recovery of constituents following treatment of LPG-I do not rule out the occurrence of small amounts of other tri- or tetra-saccharide chains. 相似文献
The structures of the peracetylated derivatives of the following alditols obtained from oligosaccharides of human milk have been established by two-dimensional, J-resolved and J-correlated, 1H-n.m.r. spectroscopy at 360 MHz: β- d-Galp-(1→3)-β- d-GlcpNAc-(1→3)-β- d-Galp-(1→4)- d-Glc-ol, α- l-Fucp-(1→2)-β- d-Galp-(1→3)-β- d-GlcpNAc-(1→3)-β- d-Galp-(1→4)- d-Glc-ol, and β- d-Galp-(1→3)-β- d-GlcpNAc-(1→3)-[β- d-Galp-(1→4)-β- d-GlcpNAc-(1→6)]-β- d-Galp-(1→4)- d-Glc-ol. 相似文献
The plant gum isolated from sap of the lac tree, Rhus vernicifera (China), was separated into two fractions having mol. wt. 84,000 and 27,700 by aqueous-phase gel-permeation chromatography. The fractions contain d-galactose (65 mol%), 4-O-methyl-d-glucuronic acid (24 mol%), d-glucuronic acid (3 mol%), l-arabinose (4 mol%), and l-rhamnose (3 mol%). Smith degradation of the carboxyl-reduced polysaccharides gives products of halved molecular weight, and these consist of a β-(1→3)-linked galactopyranan main chain and side chains made up of galactopyranose residues. Peripheral groups, such as α-d-Galp-, α-d-Galp-(1→6)-β-d-Galp-, 4-O-methyl-β-d-GlcpA-, and 4-O-methyl-β-d-GlcpA-(1→6)-β-d-Galp-, are attached to this interior core through β-(1→3)- or β-(1→6)-linkages. 相似文献
The asparagine-linked sugar chains of human milk galactosyltansferase were quantitatively released as oligosaccharides from the polypeptide backbone by hydrazinolysis. They were converted into radioactive oligosaccharides by sodium borotritiate reduction after N-acetylation, and fractionated by paper electrophoresis and by Bio-Gel P-4 column chromatography after sialidase treatment. Structural studies of each oligosaccharides by sequential exoglycosidase digestion and methylation analysis indicated that the galactosyltransferase contains bi, tri-, and probably tetra-antennary, complex-type oligosaccharides having α-d-Manp-(1→3)-[α-d-Manp-(1→6)]-β-d-Manp-(1→4)-β-d-GlcpNAc-(1→4)-α-d-[Fucp-(1→6)]-d- GlcNAc as their common core. Variation is produced by the different locations and numbers of the five different outer chains: β-d-Galp-(1→4)-d-GlcNAc, α-l-Fucp-(1→3)-[β-d-Galp-(1→4)]-d-GlcNAc, α-NeuAc-(2→6)-β-d-Galp-(1→4)-d-GlcNAc, α-l-Fucp-(1→3)-[β-d-Galp-(1→4)]-β-d-GlcpNAc-(1→3)-β-d-Galp-(1→4)-[α-l-Fucp-(1→3)]-d- GlcNAc, and α-NeuAc-(2→6)-β-d-Galp-(1→4)-β-d-GlcpNAc-(1→3)-β-d-Galp-(1→4)-[α-l-Fucp-(1→3)-β-d-GlcNAc. 相似文献
An arabinoxylan isolated from the bark of Cinnamomum zeylanicum was composed of l-arabinose and d-xylose in the molar ratio 1.6:1.0. Partial hydrolysis furnished oligosaccharides which were characterised as α-d-Xylp-(1→3)-d-Ara, β-dXylp-(1→4)-d-Xyl, β-d-Xylp-(1→4)-β-d-Xylp-(1→4)-d-Xyl, β-d-Xylp-(1→4)-β-d-Xylp-(1→4)-β-d-Xylp-Xylp-(1→4)-d-Xyl, xylopentaose, and xylohexaose. Mild acid hydrolysis of the arabinoxylan gave a degraded polysaccharide consisting of l-arabinose (8%) and d-xyolse (92%). Methylation analysis indicated the degraded polysaccharide to be a linear (1→4)-linked d-xlan in which some xylopyranosyl residues were substituted at O-2 or O-3 with l-arabinofuranosyl groups. These data together with the results of methylation analysis and periodate oxidation of the arabinoxylan suggested that it contained a (1→4)-linked β-d-xylan backbone in which each xylopyranosyl residue was substituted both at O-2 and O-3 with l-arabinofuranosyl, 3-O-α-d-xylopyranosyl-l-arabinofuranosyl, and 3-O-l-arabinofuranosyl-l-arabinofuranosyl groups. 相似文献
Horse B-active and non-B-active glycoproteins from gastric mucosae are indistinguishable in their precipitating abilities with concanavalin A, anti-BP1, type XIV horse antipneumococcal serum, the lectin from Lotus tetragonolobus and a group 1 anti-I serum, Ma; no Lea or Leb activity was found. Each was subjected to catalyzed release of its oligosaccharide chains by 0.05 n NaOH in 1 m NaBH4. Destruction of serine, threonine and 2-acetamido-2-deoxygalactopyranose (dGalNAc) was associated with production of alanine, α-aminobutyric acid and N-acetyl-d-galactosaminitol, as expected for a carbohydrate to peptide linkage via dGalNAc to serine or threonine. No evidence of basecatalyzed peeling was seen. Bio-Gel P-2 elution patterns of the salt-free oligosaccharides from the two preparations were compared. Unlike results obtained with human ovarian cyst substances, very little material was excluded. The largest-size chains are in the range of deca- or dodecasaccharides, and a reduced octasaccharide was isolated. The four most abundant amino acids in both B-active and non-B-active materials are threonine, serine, proline and glutamic acid, which together account for 60% of the weight of amino acids. 相似文献
α-Amylases have been found to convert starch and glycogen, in part, to products other than hemiacetal-bearing entities (maltose, maltodextrins, etc.)—hitherto, the only products obtained from natural α-glucans by α-amylolysis. Glycosides of maltosaccharides were synthesized by purified α-amylases acting on starch or bacterial glycogen in the presence of p-nitrophenyl α- or β-d-glucoside. From a digest with crystallized B. subtilis var. amyloliquefaciens α-amylase, containing 4 mg/ml of [14C]glycogen and 40 mmp-NP β-d-glucoside, three pairs of correspondingly labeled glycosides and sugars were recovered: p-NP α-d-[14C]glucopyranosyl (1 → 4) β-d-glucopyranoside, and [14C]glucose; p-NP α-[14C]maltosyl (1 → 4) β-d-glucopyranoside, and [14C]maltose; p-NP α-[14C]maltotriosyl (1 → 4) β-d-glucopyranoside, and [14C]maltotriose. The three glycosides accounted for 11.4% of the [14C]glycogen donor substrate; the three comparable sugars, for 30.4%; higher maltodextrins, for 58.2%. Calculations based on the molar yields of all reaction products show that [14C]glycosyl moieties were transferred from donor to p-NP β-d-glucoside with a frequency of 0.234 relative to all transfers to water. This is a very high value considering the minute molar ratio (0.0007) of β-d-glucoside-to-water concentration. Less striking but similar findings were obtained with cryst. hog pancreatic and Aspergillus oryzae α-amylases. The results extend earlier findings (Hehre et al., Advan. Chem. Ser. (1973) 117, 309) in showing that α-amylases have a substantial capacity to utilize the C4-carbinols of certain d-glucosyl compounds as acceptor sites. 相似文献
Syntheses, based on silver trifluoromethanesulfonate-promoted Koenigs-Knorr type condensations, are described of the d-glucotrioses, β-d-Glcp-(1→3)-β-d-Glcp-(1→4)-d-Glcp and β-d-Glcp-(1→4)-β-d-Glcp-(1→3)-d-Glcp, and the d-Glucotetraoses, β-d-Glcp-(1→3)-β-d-Glcp-(1→4)-β-d-Glcp-(1→4)-d-Glcp, β-d-Glcp-(1→4)-β-d-Glcp-(1→3)-β-d-Glcp-(1→4)-d-Glcp, and β-d-Glcp-(1→4)-β-d-Glcp-(1→4)-β-d-Glcp-(1→3)-d-Glcp, corresponding to the tri- and tetra-saccharide units in the linear chains of (1→4)- and (1→3)-linked β-d-glucopyranosyl residues of lichenan, and of oat and barley β-d-glucans. 相似文献