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1.
Sylvain Bourgerie Yannis Karamanos Sylvie Berger Raymond Julien 《Glycoconjugate journal》1992,9(4):162-167
Peptide-N
4-(N-acetyl--glucosaminyl) asparagine amidase F (PNGase F) and endo--N-acetyl glucosaminidase F (Endo F) activities were monitored during cultivation ofFlavobacterium meningosepticum using a new fluorescence-HPLC procedure based on a commercially available substrate. The PNGase F activity reached a maximum level at the end of the log phase and remained constant during the stationary phase, while Endo F continuously increased until late stationary phase. PNGase F obtained at the end of the log phase was less contaminated by other proteins compared with late stationary phase.Abbreviations Con A
concanavalin A
- Endo F
endo--N-acetyl glucosaminidase F (EC 3.2.1.96)
- GlcNAc
N-acetylglucosamine
- PNGase F
peptide-N
4-(N-acetyl--glucosaminyl) asparagine amidase F (EC 3.5.1.52). 相似文献
2.
Jan B L Damm Hans Voshol Karl Hård Johannis P Kamerling Gijs W K Van Dedem Johannes F G Vliegenthart 《Glycoconjugate journal》1988,5(3):221-233
TheN-linked carbohydrate chains of the-subunit of highly purified urinary human chorionic gonadotropin have been re-investigated. The oligosaccharides were released enzymatically by peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase-F, and fractionated by a combination of FPLC and HPLC. As a result of the application of improved fractionation methods, apart from the earlier reported carbohydrate chains, also small amounts of trisialo tri- and tri-antennary oligosaccharides were found. The primary structures of the latter carbohydrate chains have been determined by 500-MHz1H-NMR spectroscopy to beAbbreviations hCG
human chorionic gonadotropin
- hCG-
-subunit
- hCG-
-subunit
- PNGase-F
peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase-F (E.C. 3.5.1.52)
- endo-F
endo--N-acetylglucosaminidase-F (E.C. 3.2.1.96)
- SDS
sodium dodecyl sulphate
- PAGE
polyacrylamide gel electrophoresis
- CBB
coomassie brilliant blue R 250
- GlcNAc
N-acetylglucosamine
- NeuAc
N-acetylneuraminic acid
- Man
mannose
- Gal
galactose
- Fuc
fucose 相似文献
3.
Gabriele D'andrea Jan B Bouwstra Johannis P Kamerling Johannes F G Vliegenthart 《Glycoconjugate journal》1988,5(2):151-157
Ascorbic acid oxidase (E.C.1.10.3.3) from the green zucchini squash (Cucurbita pepo medullosa) is a copper-containing glycoprotein which catalyzes the reaction:l-ascorbic acid +1/2 O2l-dehydroascorbic acid + H2O. The carbohydrate content of the purified plant glycoprotein amounted to 3% (w/w), and monosaccharide analysis revealed the carbohydrate moiety to be of theN-glycosidic type. The carbohydrate chains were released from the apoenzyme by digestion with PNGase-F immobilized on Sepharose 4B. After fractionation on Bio-Gel P-2 and purification on Mono-Q, the neutral oligosaccharide was investigated by 500-MHz1H-NMR spectroscopy. The primary structure of theN-linked carbohydrate chain was established to be:
Abbreviations AAO
ascorbic acid oxidase
- PNGase-F
peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase-F
- GalNAc
N-acetylgalactosamine
- GlcNAc
N-acetylglucosamine
- Man
mannose
- Xyl
xylose
- GLC
gas-liquid chromatography
- FPLC
fast protein liquid chromatography
- NMR
nuclear magnetic resonance
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis 相似文献
4.
Sabine Lhernould Yannis Karamanos Sylvain Bourgerie Gerard Strecker Raymond Julien Henri Morvan 《Glycoconjugate journal》1992,9(4):191-197
We have previously isolated mannoside and xylomannoside oligosaccharides with one or two terminal reducingN-acetylglucosamine residues from the extracellular medium of white campion (Silene alba) suspension culture. We have now demonstrated the presence of peptide-N
4-(N-acetylglucosaminyl)asparagine amidase (PNGase) activity in cell extracts as well in the culture medium that could explain the production of those compounds. An additional xylomannoside, (GlcNAc)Man3(Xyl)GlcNAc(Fuc)GlcNAc, was characterized, and1H- and13C-NMR assignments for the oligosaccharide Man3(Xyl)GlcNAc(Fuc)GlcNAc were obtained using homonuclear and heteronuclear spectroscopy (COSY).Abbreviations Endo
endo--N-acetylglucosaminidase
- Fuc
fucose
- GlcNAc
N-acetylglucosamine
- Man
mannose
- NMR
nuclear magnetic resonance
- PNGase
peptide-N
4-(N-acetylglucosaminyl)asparagine amidase
- Xyl
xylose 相似文献
5.
Jan B L Damm Johannis P Kamerling Gijs W K van Dedem Johannes F G Vliegenthart 《Glycoconjugate journal》1987,4(2):129-144
For the structural analysis of the carbohydrate chains ofN-,O-glycoproteins a straightforward strategy was developed based on the cleavage of theN-linked chains with immobilized peptide-N
4-(N-acetyl--glucosaminyl) asparagine amidase-F (PN-Gase-F) fromFlavobacterium meningosepticum, followed by alkaline borohydride treatment of the remainingO-glycoprotein material. This methodology was applied to the isolation of the Asn- and Ser-linked carbohydrate chains of human chorionic gonadotrophin. The structures of the isolated oligosaccharides were verified by 500-MHz1H-NMR spectroscopy. The Asn-linked sugar chains were shown to be: NeuAc2-3Gal1-4GlcNAc1-2Man1-6[NeuAc2-3Gal1-4GlcNAc1-2Man1-3]Man 1-4GlcNAc1-4[Fuc1-6]0-1GlcNAc and Man1-6[NeuAc2-3Gal1-4GlcNAc1-2Man 1-3]Man1-4GlcNAc1-4GlcNAc. Also some minor constituents occurred. The structures of the Ser-linked oligosaccharides were established in the form of their oligosaccharide-alditols as: NeuAc2-3Gal1-3[NeuAc2-6]GalNAc, NeuAc2-3Gal 1-3GalNAc and NeuAc2-3Gal1-3[NeuAc2-3Gal1-4GlcNAc1-6]GalNAc.Abbreviations hCG
human chorionic gonadotrophin
- hCG-
-subunit
- hCG-
-subunit
- ElA
enzyme immunoassay
- PNGase-F
peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase-F (EC 3.5.1.52)
- SDS
sodium dodecyl sulphate
- GalNAc
N-acetylgalactosamine
- GlcNAc
N-acetylglucosamine
- NeuAc
N-acetylneuraminic acid
- Man
mannose
- Gal
galactose
- Fuc
fucose 相似文献
6.
Normal rat kidney cells, non-productively infected with the anaemia-inducing variant of Friend spleen focus-forming virus (F-SFFVA), were metabolically labelled with [2-3H]mannose. The primary translation product of the viral envelope gene (env), representing a glycoprotein with an apparent molecularM
r of 55 000 (gp55), was isolated from cell lysates by immunoaffinity chromatography and purified by preparative SDS/PAGE. Radiolabelled oligosaccharides, released from tryptic glycopeptides by treatment with endo--N-acetylglucosaminidase H, were characterized chromatographically, by enzymic digestion and by acetolysis. The results revealed that F-SFFVA gp55 obtained from this source carried predominantly oligomannose type sugar chains with five to nine mannoses. As a characteristic feature, glycans with seven to nine mannoses contained, in part, an additional glucose residue. Although the amount of glucosylated species found was higher in F-SFFVA gp55 (about 25% of total endo-H-sensitive oligosaccharides) than in gp55 of the corresponding polycythaemia-inducing variant (F-SFFVP, 16.3%), the overall glycosylation pattern of the F-SFFVA
env product closely resembled that of F-SFFVP gp55 [Strubeet al. (1988)J Biol Chem
263:3762–71]. Hence, our results demonstrate that the different intracellular processing and transport of the primary F-SFFVA
env product cannot be attributed to aberrant trimming of its oligomannose type glycans.Abbreviations endo H
endo--N-acetylglucosaminidase H fromStreptomyces griseus
-
env
envelope gene
- Env protein
translation product ofenv
- F-SFFV
Friend spleen focus-forming virus
- F-SFFVA
anaemia-inducing variant of F-SFFV
- F-SFFVP
polycythaemia-inducing variant of F-SFFV
- Hex
hexose
- NRK
normal rat kidney
- PNGase F
peptide-N
4-(N-acetyl--glucosaminyl)asparagine amidase F fromFlavobacterium meningosepticum 相似文献
7.
Peptide-N4-(N-acetyl--glucosaminyl)asparagine amidase F (PNGase F) fromFlavobacterium meningosepticum is a highly useful enzyme for the structural analysis of N(asparagine)-linked carbohydrate chains derived from glycoproteins. The enzyme was enriched using a published procedure [Tarentino AL, Gomez CM, Plummer TH, Jr (1984) Biochemistry 1985:4665–71; Tarentino AL, Plummer TH, Jr (1987) Methods Enzymol 138:770–78] and further purified by hydrophobic interaction HPLC on a weak hydrophobic TSK-Ether column from which it was eluted by a decreasing gradient of 1.7 M ammonium sulphate in 100 mM sodium phosphate, pH 7.0, containing 5 mM EDTA.To determine the optimal conditions for a complete deglycosylation of glycoproteins by PNGase F, experiments were performed with human
1-acid glycoprotein, because the five complex type carbohydrate chains are quite resistant to enzymic hydrolysis. The influence of different detergents on the enzyme reaction was studied. Complete deglycosylation of human
1-acid glycoprotein was achieved by the use of 60 mU/ml PNGase F in 0.25 M sodium phosphate buffer, pH 8.6, containing 0.2% (w/v) SDS, 20 mM mercaptoethanol and 0.5% Mega-10. 相似文献
8.
Sabine Lhernould Yannis Karamanos Patrice Lerouge Henri Morvan 《Glycoconjugate journal》1995,12(1):94-98
The peptide-N
4-(N-acetylglucosaminyl) asparagine amidase (PNGase Se) earlier described [Lhernould S., Karamanos Y., Bourgerie S., Strecker G., Julien R., Morvan H. (1992)Glycoconjugate J
9:191–97] was partially purified from culturedSilene alba cells using affinity chromatography. The enzyme is active between pH 3.0 and 6.5, and is stable in the presence of moderate concentrations of several other protein unfolding chemicals, but is readily inactivated by SDS. Although the enzyme cleaves the carbohydrate from a variety of animal and plant glycopeptides, it does not hydrolyse the carbohydrate from most of the corresponding unfolded glycoproteins in otherwise comparable conditions. The substrate specificity of this plant PNGase supports the hypothesis that this enzyme could be at the origin of the production of unconjugated N-glycans in a suspension medium of culturedSilene alba cells.Abbreviations GlcNAc
N-acetylglucosamine
- PNGase
peptide-N
4-(N-acetylglucosaminyl) asparagine amidase
- BSA
bovine serum albumin
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- TLC
thin layer chromatography
- HPAEC-PAD
High pH anion exchange chromatography-pulsed amperometric detection 相似文献
9.
Cytidine-5-monophospho-N-acetylneuraminic acid:-galactoside 2-6sialyltransferase was purified from bovine colostrum by two sequential affinity chromatography steps on CDP-ethanolamine-Sepharose and CDP-ethanolamine-(N-caproylamino-)-Sepharose, respectively. While the conditions for elution were those of Paulsonet al. [J Biol Chem (1977) 252:3256–62], the ligand of the second affinity column was coupled to Sepharose by using 6-aminocaproic acid as linker. The ease of this procedure allows rapid synthesis of bulk quantities of ligand.Highly purified preparations of sialyltransferase were obtained which moved on gradient gel electrophoresis as a single band of 76 kDa and on dodecylsulphate electrophoresis as a single band of 54 kDa. The product of the reaction between lactose and CMP-N-acetylneuraminic acid catalyzed by the purified sialyltransferase was identified by high-resolution 500 MHz1H-NMR spectroscopy as Neu5Ac2-6Gal1-4Glc. 相似文献
10.
J A Van Kuik R A Hoffmann J H G M Mutsaers H Van Halbeek J P Kamerling J F G Vliegenthart 《Glycoconjugate journal》1986,3(1):27-34
The 500-MHz1H-NMR characteristics of theN-linked carbohydrate chain Man1-6[Xyl1-2]Man1-4GlcNAc1-4[Fuc1-3]GlcNAc1-NAsn of the proteolytic enzyme bromelain (EC 3.4.22.4) from pineapple stem were determined for the oligosaccharide-alditol and the glycopeptide, obtained by hydrazinolysis and Pronase digestion, respectively. The1H-NMR structural-reporter-groups of the (1–3)-linked fucose residue form unique sets of data for the alditol as well as for the glycopeptide. 相似文献
11.
Summary
Candida pelliculosa var. acetaetherius is a strain of yeast which can utilize cellobiose as the carbon source. From a gene library prepared from this yeast, the -glucosidase gene has been cloned in a S. cerevisiae host using a chromogenic substrate, 5-bromo-4-chloro-3-indolyl--glucoside as an indicator. It was proved by Southern analysis that the DNA fragment carrying the -glucosidase gene originated from C. pelliculosa. -Glucosidase produced by S. cerevisiae transformants was secreted into the periplasmic space. In Candida, -glucosidase was not induced by cellobiose but was derepressed by lowering the concentration of glucose. The regulation of -glucosidase synthesis in S. cerevisiae carrying the cloned -glucosidase was not clear compared with that in Candida, however, the enzyme activity in low glucose medium (0.05%) was reproducibly higher than in high glucose medium (2%). We have found the sequence that controls the expression of the -glucosidase gene negatively in S. cerevisiae. 相似文献
12.
R. M. Kondratenko L. A. Baltina S. R. Mustafina E. V. Vasil'eva R. Pompei D. Deidda O. A. Plyasunova A. G. Pokrovskii G. A. Tolstikov 《Russian Journal of Bioorganic Chemistry》2004,30(3):275-282
Glycyrrhizic acid and its 30-methyl ester were conjugated with 2-amino-1,3,4,6-tetra-O-acetyl-2-deoxy--D-glucopyranose, 2,3,4,6-tetra-O-acetyl--D-glucopyranosyl amine, 2,3,4-tri-O-acetyl--L-arabinopyranosyl amine, 2-acetamido-2-deoxy--D-glucopyranosyl amine, and -D-galactopyranosyl amine using N,N-dicyclohexylcarbodiimide and its mixtures with N-hydroxybenzotriazole. Structures of the conjugates were confirmed by IR, UV, 1H, and 13C NMR spectroscopy. The glycoconjugate with the residues of 2-acetamido-2-deoxy--D-glucopyranosyl amine in the carbohydrate part of its molecule exhibited antiviral activity (ID50 4 g/ml) toward the herpes simplex type 1 virus (HSV-1) in the VERO cell culture. Two compounds demonstrated anti-HIV-1 activity (50–70% inhibition of p24) in a culture of MT-4 cells at concentrations of 0.5–20 g/ml. 相似文献
13.
Summary The principal pancreatic islets of the teleost Scorpaena scropha are found ultrastructurally to contain four different kinds of parenchymal cells, viz.
1-(= D),
2-, -and agranular cells. The -cells show considerable variations in the shape of the secretory granules. A peculiar feature is that many of these granules are composed of fibrillar subunits, often in parallel arrangement. All -granules are surrounded by membranes and between the membrane and the granule core there is a moderately wide electron lucent space. The electron density of the cytoplasm in the -cells varies somewhat. The
2-cells possess typical secretory granules with an electron dense core and a closely applied membrane. The secretory granules in the
1-cells show also a closely applied membrane but a less dense core. Also in the -cells the electron opacity of the cytoplasm varies. The agranular cells are mainly characterized by low cytoplasmic electron density, narrow cisterns of endoplasmic reticulum and sometimes a laminated Golgi complex. Small immature secretory granules are occasionally seen in the cytoplasm of these cells. The significance of the fibrillar -granules remains obscure.This work was supported by grants from the Nordic Insulin Fund, the Town of Umeå, the Swedish Medical Research Council (Project No. B69-12X-718-04A), and by a postdoctoral fellowship from the United States Public Health Service. 相似文献
14.
Internodal cells of the giant alga Chara corallina were perfused internally to replace the native cytoplasm, tonoplast and vacuole with artificial cytoplasm. Sodium efflux from perfused cells, measured by including 22Na in the perfusion media, was increased by increasing the internal sodium concentration and by decreasing the external pH, and was inhibited by external application of the renal diuretic amiloride. The sodium efflux was markedly ATP-dependent, with a 50-fold decrease in efflux observed after perfusion with media lacking ATP. Efflux in the presence of ATP was reduced by 33% by inclusion of 10 M N,N-dicyclohexylcarbodiimide in the perfusion medium. The membrane potential of the perfused cells approximated that of intact cells from the same culture. It is suggested that sodium efflux in perfused Chara cells proceeds via a secondary antiporter with protons, regulated by ATP in a catalytic role and with the proton motive force acting as the energy source.Abbreviations DCCD
N,N-dicyclohexylcarbodiimide
- EGTA
ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid
- Mes
2-(N-morpholino)ethanesulphonic acid
- Mops
3(N-morpholino)propanesulphonic acid
- Taps
tris(hydroxymethyl)methylaminopropanesulphonic acid 相似文献
15.
Herman Van Halbeek Johannes F G Vliegenthart Hitoo Iwase Suchen Li Yu-Teh Li 《Glycoconjugate journal》1985,2(3-4):235-253
Dansyl glyco-asparagines were prepared from a partially fractionated mixture of hen egg white glycoproteins. Reverse-phase high performance liquid chromatography (HPLC) on a silica-based octadecyl column yielded ten such derivatives in a virtually pure state. The detailed structures of the glyco-asparagines were identified by 500-MHz1H nuclear magnetic resonance (NMR) spectroscopy. Two of them were found to be of the oligomannosideN-type, four were of the intersected-hybridN-type and another four were of the intersected multi-antennaryN-type. In monogalactosylated, intersected structures the galactose residue was proved by1H-NMR to be attached in (1–4)-linkage to the GlcNAc1-4Man1–3 branch.Dansyl glyco-asparagines turned out to be suitable derivatives for1H-NMR spectroscopic analysis. The combination of HPLC and high-resolution NMR spectroscopy of such derivatives proved to be a powerful technique in studying the (micro-)heterogeneity of sugar chains in glycoproteins.Abbreviations dns
dansyl (5-dimethylaminonaphthalene-1-sulfonyl)
- ODS
octadecyl-silica
- WEFT
water-eliminating Fourier transform
- DSS
sodium 4,4-dimethyl-4-silapentane-1-sulfonate
- OVA
ovalbumin
- OVM
ovomucoid
- OVT
ovotransferrin 相似文献
16.
Rudolf Geyer Silvia Diabaté Hildegard Geyer Hans-Dieter Klenk Heiner Niemann Stephan Stirm 《Glycoconjugate journal》1987,4(1):17-32
Fowl plague virus, strain Dutch, was metabolically labeled withd-[2-3H]mannose, or withd-[6-3H]glucosamine, and the small subunit (HA2; 0.8 mg in total) of the viral hemagglutinin was isolated by preparative sodium dodecylsulfate-polyacrylamide gel electrophoresis. After proteolytic digestion, the radioactive oligosaccharides were sequentially liberated from the glycopeptides by treatment with different endo--N-acetylglucosaminidases and with peptide:N-glycosidase or, finally, by hydrazinolysis. In this manner, four groups of glycans could be obtained by consecutive gel filtrations and were subfractionated by HPLC. The structures of the individual oligosaccharides were analyzed by micromethylation, by acetolysis or by digestion with exoglycosidases. The major species amongst the high mannose glycans at Ans-406 of the viral glycopolypeptide were found to be Man1-2Man1-3(Man1-2Man1-6)Man1-6(Man1-2Man1-2Man1-3)Man1-4GlcNac1-4GlcNAc and Man1-3(Man1-2Man1-6)Man1-6(Man1-2Man1-2Man1-3)Man1-4GlcNAc1-4GlcNAc, while the complex glycans at Asn-478 are predominantly GlcNAc1-2Man1-3(GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc (lacking, in part, one of the outerN-acetylglucosamine residues) and GlcNAc1-2Man1-3(Gal1-4GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc.Abbreviation BSA
bovine serum albumin
- endo D (F,H)
endo--N-acetyl-d-glucosaminidase D (F,H)
- HA
hemagglutinin (HA1, large subunit of HA
- HA2
small subunit
- FPV
fowl plague virus
- PNGase F
peptide:N-glycosidase F
- SDS
sodium dodecylsulfate 相似文献
17.
A simple synthesis of octyl 3,6-di-O-(-d-mannopyranosyl)--d-mannopyranoside is described. The key features of the synthetic scheme are the formation of the -mannosidic linkage by 1-O-alkylation of 2,3,4,6-tetra-O-acetyl-,-d-mannopyranose with octyl iodide and glycosylation of unprotected octyl -d-mannopyranoside using limiting acetobromomannose. The trisaccharide is shown to be an acceptor forN-acetylglucosaminyltransferase-I with aK
M of 585 µm. 相似文献
18.
Recently, we have reported purification and characterization of a de-N-glycosylating enzyme, peptide:N-glycanase (PNGase) found in C3H mouse fibroblast L-929 cells, and designated L-929 PNGase [Suzuki T, Seko A, Kitajima K, Inoue Y, Inoue S (1994)J Biol Chem
269, 17611–18]. The unique properties of L-929 PNGase are that the enzyme had a high affinity to the substrate glycopeptide (e.g.K
m=114 µm for fetuin derived glycopentapeptide) and that the PNGase-catalysed reaction is strongly inhibited by the released free oligosaccharides but not by the free peptides formed, suggesting that L-929 PNGase is able to bind to a certain type of carbohydrate chain. In this study, we report the new findings of the mannan-binding property of L-929 PNGase; the de-N-glycosylating enzyme activity of L-929 PNGase was inhibited by yeast mannan and triomannose, Man1 3(Man1 6)Man, but not by mannose and -methyl-d-mannoside. Furthermore, L-929 PNGase was revealed to bind to the glycan moiety of yeast mannan by using mannan-conjugated Sepharose 4B gel as a ligand, suggesting that L-929 PNGase could serve not only as an enzyme but also as a carbohydrate recognition proteinin vivo. Such dual properties found for animal-derived L-929 PNGase are unique and are not shared with other previously characterized plant- and bacterial-origin PNGases — PNGase A and PNGase F, respectively.Abbreviations GLC
gas liquid chromatography
- GlcNAc-Asn
2-acetamido-1--(l-aspartamido)-1,2-dideoxy-d-glucose
- BSA
bovine serum albumin
- DTT
dithiothreitol
- EDTA
ethylenediaminetetraacetic acid
- Gal
d-galactose
- GlcNAc
N-acetyl-d-glucosamine
- Man
d-mannose; triomannose, Man1 3(Man1 6)Man;
- MES
2-(N-morphorino)ethanesulfonic acid
- NeuAc
N-acetyl-neuraminic acid
- PNGase
peptide:N
4-(N-accetyl-glucosaminyl)asparagine amidase (peptide:N-glycanase,EC 3.5.1.52)
- PNP
p-nitrophenyl 相似文献
19.
William Kuhns Volker Rutz Hans Paulsen Khushi L. Matta Michael A. Baker Marijke Barner Maria Granovsky Inka Brockhausen 《Glycoconjugate journal》1993,10(5):381-394
To elucidate control mechanisms ofO-glycan biosynthesis in leukemia and to develop biosynthetic inhibitors we have characterized core 2 UDP-GlcNAc:Gal1-3GalNAc-R(GlcNAc to GalNAc) 6-N-acetylglucosaminyl-transferase (EC 2.4.1.102; core 2 6-GlcNAc-T) and CMP-sialic acid: Gal1-3GalNAc-R 3-sialyltransferase (EC 2.4.99.4; 3-SA-T), two enzymes that are significantly increased in patients with chronic myelogenous leukemia (CML) and acute myeloid leukemia (AML). We observed distinct tissue-specific kinetic differences for the core 2 6-GlcNAc-T activity; core 2 6-GlcNAc-T from mucin secreting tissue (named core 2 6-GlcNAc-T M) is accompanied by activities that synthesize core 4 [GlcNAc1-6(GlcNAc1-3)GalNAc-R] and blood group I [GlcNAc1-6(GlcNAc1-3)Gal-R] branches; core 2 6-GlcNAc-T in leukemic cells (named core 2 -GlcNAc-T L) is not accompanied by these two activities and has a more restricted specificity. Core 2 6-GlcNAc-T M and L both have an absolute requirement for the 4- and 6-hydroxyls ofN-acetylgalactosamine and the 6-hydroxyl of galactose of the Gal1-3GalNAc-benzyl substrate but the recognition of other substituents of the sugar rings varies, depending on the tissue. 3-sialytransferase from human placenta and from AML cells also showed distinct specificity differences, although the enzymes from both tissues have an absolute requirement for the 3-hydroxyl of the galactose residue of Gal1-3GalNAc-Bn. Gal1-3(6-deoxy)GalNAc-Bn and 3-deoxy-Gal1-3GalNAc-Bn competitively inhibited core 2 6-GlcNAc-T and 3-sialyltransferase activities, respectively.Abbreviations AFGP
antifreeze glycoprotein
- AML
acute myeloid leukemia
- Bn
benzyl
- CML
chronic myelogenous leukemia
- Fuc
l-fucose
- Gal, G
d-galactose
- GalNAc, GA
N-acetyl-d-galactosamine
- GlcNAc, Gn
N-acetyl-d-glucosamine
- HC
human colonic homogenate
- HO
hen oviduct microsomes
- HPLC
high performance liquid chromatography
- mco
8-methoxycarbonyl-octy
- Me
methyl
- MES
2-(N-morpholino)ethanesulfonate
- MK
mouse kidney homogenate
- onp
o-nitrophenyl
- PG
pig gastric mucosal microsomes
- pnp
p-nitrophenyl
- RC
rat colonic mucosal microsomes
- SA
sialic acid
- T
transferase
Enzymes: UDP-GlcNAc:Gal1-3GalNAc-R (GlcNAc to GalNAc) 6-N-acetylglucosaminyltransferase,O-glycan core 2 6-GlcNAc-transferase, EC 2.4.1.102; CMP-sialic acid: Gal1-3GalNAc-R 3-sialyltransferase,O-glycan 3-sialic acid-transferase, EC 2.4.99.4. 相似文献
20.
Alf Gunnarsson 《Glycoconjugate journal》1987,4(3):239-245
O-Methylation of simple neutral oligosaccharides is readily accomplished in dimethyl sulphoxide containing solid sodium hydroxide and methyl iodide [Cincanu I, Kerek F (1984) Carbohydr Res 131209-17]. This procedure has been extended to 2-acetamido-2-deoxy sugars and sialic acid-containing oligosaccharides. CompleteO-andN-methylation was in most cases achieved in 15 min. Esterification of carboxylic groups in uronic acids was fast and resulted in concomitant -elimination. The method is also suitable for methylation of glycoproteins and glycosphingolipids. Polysaccharides can also be methylated by this technique. Analysis of the products by gas-liquid chromatography and mass spectrometry showed no degradation products.Abbreviations lacto-N-tetraose
LcOse4, Gal3GlcNAc3Gal4Glc
- lacto-N-fucopentaose III
III3Fuc-nLcOse4, Gal4[Fuc3]GlcNAc3Gal4Glc
- trihexosylceramide
GbOse3Cer, Gal4Gal4Glc1-1Cer
- globoside
GbOse4Cer, GalNAc3Gal4Glc1-1Cer
- FAB-MS
fas atom bombardment mass spectrometry 相似文献