共查询到20条相似文献,搜索用时 31 毫秒
1.
We have developed a new method for the large scale preparation of pyridylaminated (PA-) oligosaccharides from glycoproteins. Phenol/chloroform extration was adapted for the removal of protein and excess 2-aminopyridine, improving the efficiency of preparation. From a 2.5 g sample of human apo-transferrin, 25–30 mol of agalacto biantennary PA-oligosaccharide could be obtained. By increasing the concentration of PA-oligosaccharide substrate, we were able to detect a very low level ofN-acetylglucosaminlytransferase IV activity in CHO cell extracts.Abbreviations PA
2-aminopyridine
- SDS
sodium dodecyl sulfate
- GlcNAc
N-acetylglucosamine
- GnT
N-acetylglucosaminyltransferase
- Gn,Gn-bi-PA
GlcNAc1-2Man1-3(GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc-2-aminopyridine
- Gn,Gn,Gn-tri-PA
GlcNAc1-2(GlcNAc1-4)Man1-3(GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc-2-aminopyridine
- Gn,Gn,Gn-trí-PA
GlcNAc1-2Man1-3({GlcNAc1-2(GlcNAc1-6)Man1-6})Man1-4GlcNac1-4GlcNAc-2-aminopyridine
- Gn,(Gn),Gn-bi-PA
GlcNAc1-2Man1-3(GlcNAc1-4)(GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc-2-aminopyridine 相似文献
2.
Erika Staudacher Thomas Dalik Petra Wawra Friedrich Altmann Leopold März 《Glycoconjugate journal》1995,12(6):780-786
An 1,3-fucosyltransferase was purified 3000-fold from mung bean seedlings by chromatography on DE 52 cellulose and Affigel Blue, by chromatofocusing, gelfiltration and affinity chromatography resulting in an apparently homogenous protein of about 65 kDa on SDS-PAGE. The enzyme transferred fucose from GDP-fucose to the Asn-linkedN-acetylglucosaminyl residue of an N-glycan, forming an 1,3-linkage. The enzyme acted upon N-glycopeptides and related oligosaccharides with the glycan structure GlcNAc2Man3 GlcNAc2. Fucose in 1,6-linkage to the asparagine-linked GlcNAc had no effect on the activity. No transfer to N-glycans was observed when the terminal GlcNAc residues were either absent or substituted with galactose.N-acetyllactosamine, lacto-N-biose andN-acetylchito-oligosaccharides did not function as acceptors for the 1,3-fucosyltransferase.The transferase exhibited maximal activity at pH 7.0 and a strict requirement for Mn2+ or Zn2+ ions. The enzyme's activity was moderately increased in the presence of Triton X-100. It was not affected byN-ethylmaleimide.Abbreviations 1,3-Fuc-T
GDP-fucose:-N-acetylglucosamine(Fuc to Asn-linked GlcNAc)1,3-fucosyltransferase
- 1,6-Fuc-T
GDP-fucose:-N-acetylglucosamine(Fuc to Asn-linked GlcNAc) 1,6-fucosyltransferase
- PA
pyridylamino
- GnGn
GlcNAc1-2Man1-6(GlcNAc1-2Man1-3)Man1-4GlcNAc1-4GlcNAc
- GnGnF3
GlcNAc1-2Man1-6(GlcNAc1-2Man1-3)Man1-4GlcNAc1-4(Fuc1-3)GlcNAc
- GnGnF6
GlcNAc1-2-Man1-6(GlcNAc1-2Man1-3)Man1-4GlcNAc1-4(Fuc1-6)GlcNAc
- GnGnF3F6
GlcNAc1-2Man1-6(GlcNAc1-2Man1-3)Man1-4GlcNAc1-4(Fuc1-3)[Fuc1-6]GlcNAc
- MM
Man1-6(Man1-3)Man1-4GlcNAc1-4GlcNAc
- MMF3
Man1-6(Man1-3)Man1-4GlcNAc1-4(Fuc1-3)GlcNAc
- MMF3F6
Man1-6(Man1-3)Man1-4GlcNAc1-4(Fuc1-3)[Fuc1-6]GlcNAc 相似文献
3.
When 7721 human hepatocarcinoma cells were treated with 100nm phorbol-12-myristate-13-acetate (PMA), the activity ofN-acetylglucosaminyltransferase V(GnT-V) in the cells varied in accordance with the activity of membranous protein kinase C (PKC), but not with that of cytosolic PKC. Quercetin, a non-specific inhibitor of Ser/Thr protein kinase, andd-sphingosine and staurosporine, two specific inhibitors of PKC, blocked the activation of membranous PKC and GnT-V by PMA. Among the three inhibitors, quercetin was least effective. The inhibitory rates of quercetin and staurosporine toward membranous PKC and GnT V were proportional to the concentrations of the two inhibitors. The activities of GnT V and membranous protein kinase A (PKA) were also induced in parallel by dibutyryl cAMP (db-cAMP) and this induction was blocked by a specific PKA inhibitor. When cell free preparations of 7721 cells and human kidney were treated with alkaline phosphatase (ALP) to remove the phosphate groups, the GnT V activities were decreased. These results suggest that GnT V may be activated by membranous PKC or PKA, indirectly or directly, via phosphorylation of Ser/Thr residues.Abbreviations UDP
uridine diphospho-
- GnT
N-acetylglucosaminyltransferase
- GlcNAc Gn
N-acetylglucosamine
- M
mannose
- PMA
phorbol-12-myristate-13-acetate
- PKC
protein kinase C
- PKA
protein kinase A
- cAMP
adenosine 3, 5-cyclic monophosphate
- db-cAMP
dibutyryl cAMP
- TPK
tyrosine protein kinase
- MES
2-[N-morpholino]ethanesulfonic acid
- DTT
dithiothreitol
- PMSF
phenylmethylsulfonyl fluoride
- EDTA
ethylene diamine tetraacetic acid
- EGTA
glycol-bis-(-aminoethyl) etherN,N,N,N-tetraacetic acid
- PA
2-aminopyridine
- ALP
alkaline phosphatase
- C2C2
GlcNAc1-2 Man1-6(GlcNAc1-2Man1-3)ManR
- C2,4C2
GlcNAc1-2Man1-6(GlcNAc1-4[GlcNAc1-2]Man1-3)ManR
- C2C2,6
GlcNAc1-6[GlcNAc1-2]Man1-6(GlcNAc1-2Man1-3)ManR
- C2,4C2,6
GlcNAc1-6[GlcNAc1-2]Man1-6(GlcNAc1-4[GlcNAc1-2]Man1-3)ManR where R=1-4GlcNAc1-4GlcNAcAsnX
- Gn2M3Gn2-PA
C2C2 where R=1-4GlcNAc1-4GlcNAc-PA
- Gn3M3Gn2-PA
C2C2,6 where R=1-4GlcNAc1-4GlcNAc-PA 相似文献
4.
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
5.
Structural studies of the glycopeptides of B-chain of cinnamomin – a type II ribosome-inactivating protein by nuclear magnetic resonance 总被引:1,自引:0,他引:1
Zheng Pu Yiming Li Fa-jian Hou Fahu He Naixia Zhang Houming Wu Wang-Yi Liu 《Glycoconjugate journal》2000,17(11):749-759
Cinnamomin is a plant type II ribosome-inactivating protein (RIP) isolated from the seeds of Cinnamomum camphora. It consists of two nonidentical polypeptide chains (A- and B-chain) held together through one disulfide linkage. Its A- and B-chain contain 0.3% and 3.9% sugars respectively. The B-chain of cinnamomin was digested by pronase E and then the liberated glycopeptides were separated from non-glycopeptides by gel filtration chromatography on a Bio-Gel P-4 column. Three crude glycopeptides were obtained by continuing chromatography over anion-exchange resin (AG1-X2) in the buffer of 2% pyridine-acetic acid (pH 8.3) with a polygradient elution system. Through further purification by the gel filtration chromatography and HPLC, three major glycopeptides, GP1, GP2 and GP3 were obtained. Mainly by two-dimensional Nuclear Magnetic Resonance (NMR) including TOCSY, DQF-COSY, NOESY, HMQC and HMBC, their primary structures were analyzed as: Man1,3Man1,6(Man1,3)(Xyl1,2)Man1,4GlcNAc1,4GlcNAc1-(Gly-)Asn-Asn-Thr(GP1), Man1,6(Man1,3)(Xyl1,2)Man1,4GlcNAc1,4(Fuc1,3)GlcNAc1-Asn-Ala-Thr(GP2),Man1,6(Man1,3)Man1,6(Man1,2 Man1,3)Man1,4GlcNAc1,4GlcNAc1-(Ala-)Asn-Gly-Thr(GP3). 相似文献
6.
Gabriele Möller Folkert Reck Hans Paulsen Kanwal J. Kaur Mohan Sarkar Harry Schachter Inka Brockhausen 《Glycoconjugate journal》1992,9(4):180-190
UDP-GlcNAc: Man3R 2-N-acetylglucosaminyltransferase I (GlcNAc-T I; EC 2.4.1.101) is the key enzyme in the synthesis of complex and hybrid N-glycans. Rat liver GlcNAc-T I has been purified more than 25,000-fold (M
r 42,000). TheV
max for the pure enzyme with [Man6(Man3)Man6](Man3)Man4GlcNAc4GlcNAc-Asn as substrate was 4.6 µmol min–1 mg–1. Structural analysis of the enzyme product by proton nuclear magnetic resonance spectroscopy proved that the enzyme adds anN-acetylglucosamine (GlcNAc) residue in 1–2 linkage to the Man3Man-terminus of the substrate. Several derivatives of Man6(Man3)Man-R, a substrate for the enzyme, were synthesized and tested as substrates and inhibitors. An unsubstituted equatorial 4-hydroxyl and an axial 2-hydroxyl on the -linked mannose of Man6(Man3)Man-R are essential for GlcNAc-T I activity. Elimination of the 4-hydroxyl of the 3-linked mannose (Man) of the substrate increases theK
M 20-fold. Modifications on the 6-linked mannose or on the core structure affect mainly theK
M and to a lesser degree theV
max, e.g., substitutions of the Man6 residue at the 2-position by GlcNAc or at the 3- and 6-positions by mannose lower theK
M, whereas various other substitutions at the 3-position increase theK
M slightly. Man6(Man3)4-O-methyl-Man4GlcNAc was found to be a weak inhibitor of GlcNAc-T I.Abbreviations BSA
Bovine serum albumin
- Bn
benzyl
- Fuc, F
l-fucose
- Gal, G
d-galactose
- GalNAc, GA
N-acetyl-d-galactosamine
- Glc
d-glucose
- GlcNAc, Gn
N-acetyl-d-glucosamine
- HPLC
high performance liquid chromatography
- Man, M
d-mannose
- mco
8-methoxycarbonyl-octyl, (CH2)8 COOOCH3
- Me
methyl
- MES
2-(N-morpholino)ethanesulfonate
- NMR
nuclear magnetic resonance
- PMSF
phenylmethylsulfonylfluoride
- pnp
p-nitrophenyl
- SDS
sodium dodecyl sulfate
- T
transferase
- Tal
d-talose
- Xyl
d-xylose;
- {0, 2 + F}
Man6 (GlcNAc2Man3) Man4GlcNAc4 (Fuc6) GlcNAc
- {2, 2}
GlcNAc2Man6 (GlcNAc2Man3) Man4GlcNAc4GlcNAc; M5-glycopeptide, Man6 (Man3) Man6 (Man3) Man4 GlcNAc4GlcNAc-Asn
Enzymes: GlcNAc-transferase I, EC 2.4.1.101; GlcNAc-transferase II, EC 2.4.1.143; GlcNAc-transferase III, EC 2.4.1.144; GlcNAc-transferase IV, EC 2.4.1.145; GlcNAc-transferase V, UDP-GlcNAc: GlcNAc2 Man6-R (GlcNAc to Man) 6-GlcNAc-transferase; GlcNAc-transferase VI, UDP-GlcNAc: GlcNAc6(GlcNAc2) Man6-R (GlcNAc to Man) 4-GlcNAc-transferase; Core 1 3-Gal-transferase, EC 2.4.1.122; 4-Gal-transferase, EC 2.4.1.38; 3-Gal-transferase, UDP-Gal: GlcNAc-R 3-Gal-transferase; blood group i 3-GlcNAc-transferase, EC 2.4.1.149; blood group I 6-GlcNAc-transferase, UDP-GlcNAc: GlcNAc3Gal-R (GlcNAc to Gal) 6-GlcNAc-transferase. 相似文献
7.
Inka Brockhausen Folkert Reck William Kuhns Shaheer Khan Khushi L. Matta Ernst Meinjohanns Hans Paulsen Rajan N. Shah Michael A. Baker Harry Schachter 《Glycoconjugate journal》1995,12(3):371-379
UDP-GlcNAc:GlcNAc 1-2Man1-6R (GlcNAc to Man) 1,6-N-acetylglucosaminyltransferase V (GlcNAc-T V) adds a GlcNAc1-6 branch to bi- and triantennaryN-glycans. An increase in this activity has been associated with cellular transformation, metastasis and differentiation. We have used synthetic substrate analogues to study the substrate specificity and inhibition of the partially purified enzyme from hamster kidney and of extracts from hen oviduct membranes and acute myeloid leukaemia leukocytes. All compounds with the minimum structure GlcNAc1-2Man1-6Glc/Man-R were good substrates for GlcNAc-T V. The presence of structural elements other than the minimum trisaccharide structure affected GlcNAc-T V activity without being an absolute requirement for activity. Substrates with a biantennary structure were preferred over linear fragments of biantennary structures. Kinetic analysis showed that the 3-hydroxyl of the Man1-3 residue and the 4-hydroxyl of the Man- residue of the Man1-6(Man1-3)Man-RN-glycan core are not essential for catalysis but influence substrate binding. GlcNAc1-2(4,6-di-O-methyl-)Man1-6Glc-pnp was found to be an inhibitor of GlcNAc-T V from hamster kidney, hen oviduct microsomes and acute and chronic myeloid leukaemia leukocytes.Abbreviations all
allyl
- AML
acute myeloid leukaemia
- BSA
bovine serum albumin
- CML
chronic myelogenous leukaemia
- Gal
G,d-galactose
- Glc
d-glucose
- GlcNAc
Gn,N-acetyl-d-glucosamine
- HPLC
high performance liquid chromatography
- Man
M,d-mannose
- mco
8-methoxycarbonyl-octyl, (CH2)8COOCH3
- Me
methyl
- MES
2-(N-morpholino)ethanesulfonate
- oct
octyl
- pnp
p-nitrophenyl
- T
transferase 相似文献
8.
Fumito Matsuura Masaya Ohta Khoichi Murakami Yujirou Matsuki 《Glycoconjugate journal》1993,10(3):202-213
Structures of the Asn linked oligosaccharides of quail egg-yolk immunoglobulin (IgY) were determined in this study. Asn linked oligosaccharides were cleaved from IgY by hydrazinolysis and labelled withp-aminobenzoic acid ethyl ester (ABEE) afterN-acetylation. The ABEE labelled oligosaccharides were then fractionated by a combination of Concanavalin A-agarose column chromatography and anion exchange, normal phase and reversed phase HPLC before their structures were determined by sequential exoglycosidase digestion, methylation analysis, HPLC, and 500 MHz1H-NMR spectroscopy. Quail IgY contained only neutral oligosaccharides of the following categories: the glucosylated oligomannose type (0.6%, Glc1-3Glc1-3Man9GlcNAc2; 35.6%, Glc1-3Man7–9GlcNAc2). oligomannose type (15.0%, with the structure Man5–9GlcNAc2) and biantennary complex type with core structures of-Man1-3(-Man1-6)Man1-4GlcNAc1-4GlcNAc (9.9%),-Man1-3(GlcNAc1-4)(-Man1-6)Man1-4GlcNAc1-4GlcNAc (25.1%) and-Man1-3(GlcNAc1-4)(-Man1-6)Man1-4GlcNAc1-4(Fuc1-6)GlcNAc (11.4%). Although never found in mammalian proteins, glucosylated oligosaccharides (Glc1Man7–9GlcNAc2) have been located previously in hen IgY.Abbreviations IgG, IgM, IgA, IgY
immunoglobulin G, M, A and Y, respectively
- ABEE
p-aminobenzoic acid ethyl ester 相似文献
9.
Cornelis H. Hokke Jos G. M. van der Ven Johannis P. Kamerling Johannes F. G. Vliegenthart 《Glycoconjugate journal》1993,10(1):82-90
Incubation of synthetic Man\1-4GlcNAc-OMe, GalNAc1-4GlcNAc-OMe, Glc1-4GlcNAc-OMe, and GlcNAc1-4GlcNac-OMe with CMP-Neu5Ac and rat liver Gal1-4GlcNAc (2-6)-sialyltransferase resulted in the formation of Neu5Ac2-6Man1-4GlcNAc-OMe, Neu5Ac2-6GalNAc1-4GlcNAc-OMe, Neu5Ac2-6Glc1-4GlcNAc-OMe and Neu5Ac2-6GlcNAc1-4GlcNAc-OMe, respectively. Under conditions which led to quantitative conversion of Gal1-4GlcNAc-OEt into Neu5Ac2-6Gal1-4GlcNAc-OEt, the aforementioned products were obtained in yields of 4%, 48%, 16% and 8%, respectively. HPLC on Partisil 10 SAX was used to isolate the various sialyltrisaccharides, and identification was carried out using 1- and 2-dimensional 500-MHz1H-NMR spectroscopy.Abbreviations 2D
2-dimensional
- CMP
cytidine 5-monophosphate
- CMP-Neu5Ac
cytidine 5-monophospho--N-acetylneuraminic acid
- COSY
correlation spectroscopy
- DQF
double quantum filtered
- HOHAHA
homonuclear Hartmann-Hahn
- MLEV
composite pulse devised by M. Levitt
- Neu5Ac
N-acetylneuraminic acid
- Neu5Ac2en
2-deoxy-2,3-didehydro-N-acetylneuraminic acid 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.
Poly-N-acetyllactosamines provide backbone structures for functional modifications such as sialyl Lewis X. To understand how the biosynthesis of poly-N-acetyllactosamines is regulated, two branched oligosaccharides of the structure Gal1,4GlcNAc1, 6(Gal1,4GlcNAc1,2)-Man1,6Man-octyl 1 and 2 were synthesized in which one of the terminal galactose units was selectively radiolabeled. Hexasaccharides 1 and 2 were assembled from the chemically synthesized pentasaccharide precursors GlcNAc1,6(Gal1,4GlcNAc1,2)-Man1,6Man-octyl3 and Gal1,4GlcNAc1,6(GlcNAc1, 2) - Man1,6 Man-octyl 4 respectively, through treatment with UDP-1-[3H]-Gal and 1,4 galactosyltransferase. Compounds 1 and 2 were subsequently incubated with UDP-GlcNAc and the UDP-GlcNAc: Gal1-4Glc(NAc)1,3-N-acetylglucosaminyltransferase (i-GlcNAc transferase) resulting in a partial conversion to a mixture of heptasaccharides which were purified by HPLC. The branch selectivity of the addition of N-acetylglucosamine to compounds 1 and 2 was then characterized by endo--galactosidase digestion of the heptasaccharides, followed by isolation of the resultant pentasaccharides on C18 reverse-phase silica cartridges. Comparison of the amount of radiolabel to a control reaction lacking endo--galactosidase indicated the favored site of GlcNAc addition to be the lower 1,2-branch over the 1,6-branch by a 3:1 ratio. 相似文献
12.
Ritva Niemelä Jari Natunen Elina Brotherus Annamari Saarikangas Ossi Renkonen 《Glycoconjugate journal》1995,12(1):36-44
A partially purified preparation of 1,3-fucosyltransferase(s) from human milk was used to [14C]fucosylate oligosac-charides containing Gal1-4GlcNAc units. Substitution ofN-acetyllactosamine at position 3 with a -linkedN-acetylglucosamine enhanced the reactivity of the acceptor, whereas similar substitution at position 6 was inhibitory. Thus, the trisaccharide GlcNAcl-6Gal1-4GlcNAc (5), the branched tetrasaccharide GlcNAc1-3(GlcNAc1-6)Gal1-4GlcNAc (11) and the triply branched decasaccharide GlcNAc1-3(GlcNAc1-6)Gall-4GlcNAc1-3[GlcNAc1-3(GlcNAc1-6)Gal1-4GlcNAc1-6]Gal1-4GlcNAc (26) gave remarkably poor yields of 1,3-fucosylated products in comparison to GlcNAc1-3Gal1-4GlcNAc (3). 1,4-Galactosyl derivatives of5 and11, however, gave good yields of 1,3-fucosylated products, but the fucosylation was restricted to the distalN-acetyllactosamine units of Gal1-4GlcNAc1-6Gal1-4GlcNAc (16), Gal1-4GlcNAc1-3(Gal1-4GlcNAc1-6)Gal1-4GlcNAc (18) and also in Gal1-3Gal1-4GlcNAc1-3(Gal1-3Gal1-4GlcNAc1-6)Gal1-4GlcNAc (22). Immobilized wheat germ agglutinin (WGA), possessing high affinity for16 [1], revealed no affinity for the fucosylated derivative Gal1-4(Fuc1-3)GlcNAc1-6Gal1-4GlcNAc (17). The isomeric heptasaccharides Gal1-4(Fuc1-3)GlcNAc1-3(Gal1-4GlcNAc1-6)Gal1-4GlcNAc (19) and Gal1-4GlcNAc1-3[Gal1-4(Fuc1-3)GlcNAc1-6]Gal1-4GlcNAc (20) were readily separated from each other on WGA-agarose, and so were the isomeric nonasaccharides Gal1-3Gal1-4(Fuc1-3)GlcNAc1-3(Gal1-3Gal1-4GlcNAc1-6)Gal1-4GlcNAc (23) and Gal1-3Gal1-4GlcNAc1-3[Gal1-3Gal1-4(Fuc1-3)GlcNAc1-6]Gal1-4GlcNAc (24). 相似文献
13.
Taichi Usui Masahiro Suzuki Toshinari Sato Hirokazu Kawagishi Kyoko Adachi Hiroshi Sano 《Glycoconjugate journal》1994,11(2):105-110
Transmannosylation from mannotriose (Man1-4Man1-4Man) to the 4-position at the nonreducing end N-acetylglucosaminyl residue ofN,N-diacetylchitobiose was regioselectively induced through the use of -d-mannanase fromAspergillus niger. The enzyme formed the trisaccharide Man1-4GlcNAc1-4GlcNAc (3.7% of the enzyme-catalysed net decrease ofN,N-diacetylchitobiose) from mannotriose as a donor andN,N-diacetylchitobiose as an acceptor. Mannobiose (Man1-4Man) was also shown to be useful as a donor substrate for the desired trisaccharide synthesis.Abbreviations Man
d-mannose
- (M
n) (n=1–5)
-linkedn-mer of mannose
- GlcNAc2
2-acetamido-2-deoxy--d-glucopyranosyl-(1–4)-2-acetamido-2-deoxy-d-glucose 相似文献
14.
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
15.
Hiroshi Fujimoto Megumi Isomura Tatsuo Miyazaki Ichiro Matsuo Richard Walton Tohru Sakakibara Katsumi Ajisaka 《Glycoconjugate journal》1997,14(1):75-80
GlcNAc1-2Man and GlcNAc1-6Man were synthesized using the reverse hydrolysis activity of -N-acetylglucosaminidase from both jack beans and Bacillus circulans. In turn, Gal1-4GlcNAc1-2Man and Gal1-4GlcNAc1-6Man were synthesized regioselectively using the transglycosylation activity of -galactosidase from Diplococcus pneumoniae and B. circulans, respectively. These di- and trisaccharides are important components of complex type sugar chains and will be used as intermediates in our synthetic studies. Abbreviations: pNp--GlcNAc, p-nitrophenyl 2-acetamido-2-deoxy--D-glucopyranoside; pNp--Gal, p-nitrophenyl -D-galacto-pyranoside 相似文献
16.
Endo--galactosidase (EC 3.2.1.103) ofBacteroides fragilis, at 250 mU ml–1, did not cleave the internal galactosidic linkage of the linear radiolabelled trisaccharide GlcNAc1-6Gal1-4GlcNAc, or those of the tetrasaccharides Gal1-4GlcNAc1-6Gal1-4GlcNAc and Gal1-4GlcNAc1-6Gal1-4Glc. The isomeric glycans which contained the GlcNAc1-3Gal1-4GlcNAc/Glc sequence were readily cleaved.Abbreviations GlcNAc
2-acetamido-2-deoxy-d-glucose
- Lact
lactose
- MT
maltotriose
- MTet
maltotetraose
-
R
MTet
chromatographic migration rate in relation to that of maltotetraose 相似文献
17.
Folkert Reck Matthias Springer Ernst Meinjohanns Hans Paulsen Inka Brockhausen Harry Schachter 《Glycoconjugate journal》1995,12(6):747-754
UDP-GlcNAc:Man1-3R 1-2-N-acetylglucosaminyltransferase I (GlcNAc-T I; EC 2.4.1.101) catalyses the conversion of [Man1-6(Man1-3)Man1-6][Man1-3]Man-O-R to [Man1-6(Man1-3)Man1-6] [GlcNAc1-2Man1-3]Man-O-R (R=1-4GlcNAc1-4GlcNAc-Asn-X) and thereby controls the conversion of oligomannose to complex and hybrid asparagine-linked glycans (N-glycans). GlcNAc-T I also catalyses the conversion of Man1-6(Man1-3)Man-O-octyl to Man1-6(GlcNAc1-2Man1-3)Man-O-octyl. We have therefore tested a series of synthetic analogues of Man1-6(Man1-3)Man-O-octyl as substrates and inhibitors for rat liver GlcNAc-T I. The 2-deoxy and the 3-, 4- and 6-O-methyl derivatives are all good substrates confirming previous observations that the hydroxyl groups of the Man1-6 residue do not play major roles in the binding of substrate to enzyme. In contrast, all four hydroxyl groups on the Man1-3 residue are essential since the corresponding deoxy derivatives either do not bind (2- and 3-deoxy) or bind very poorly (4- and 6-deoxy) to the enzyme. The 2- and 3-O-methyl derivatives also do not bind to the enzyme. However, the 4-O-methyl derivative is a substrate (K
m
=2.6mm) and the 6-O-methyl compound is a competitive inhibitor (K
i=0.76mm). We have therefore synthesized various 4- and 6-O-alkyl derivatives, some with reactive groups attached to anO-pentyl spacer, and tested these compounds as reversible and irreversible inhibitors of GlcNAc-T I. The 6-O-(5-iodoacetamido-pentyl) compound is a specific time dependent inhibitor of the enzyme. Four other 6-O-alkyl compounds showed competitive inhibition while the remaining compounds showed little or no binding indicating that the electronic properties of the attachedO-pentyl groups influence binding.Abbreviations GlcNAc-T I
UDP-GlcNAc:Man1-3R 1-2-N-acetylglucosaminyltransferase I (EC 2.4.1.101)
- GlcNAc-T II
UDP-GlcNAc:Man1-6R 1-2-N-acetylglucosaminyltransferase II (EC 2.4.1.143)
- MES
2-(N-morpholino)ethane sulfonic acid monohydrate 相似文献
18.
-N-Acetyl-d-hexosaminidase from Aspergillus oryzae catalysed the stereo- and regiospecific formation of the 6-O-benzylated disaccharide derivatives GalNAc1-3(6- OBn)Gal-SEt and GlcNAc1-3(6-OBn)Gal-SEt, which were obtained in transglycosylation reactions employing ethyl 6- O-benzyl-1-thio--d-galactopyranoside as acceptor. Preparative amounts of the chitobiose derivative GlcNAc1- 3GlcNAc-OPhNO2-p was prepared as well. - N-Acetyl-d-hexosaminidase from bovine testes catalysed the specific synthesis of GlcNAc1-3(6-OBn)GlcNH2-SEt and GalNAc1-3(6-OBn)GlcNH2-SEt, employing ethyl 2-amino-6-O-benzyl-2-deoxy-1-thio--d-glucopyranoside as acceptor. -d-Glucuronidase from E. coli was found to catalyse the formation of GlcA1-3(6-OBn)GlcNH2- SEt employing the same acceptor. 相似文献
19.
Kohdoh Shikata Tatsuji Yasuda Fujio Takeuchi Toshiro Konishi Munehiro Nakata Tsuguo Mizuochi 《Glycoconjugate journal》1998,15(7):683-689
In order to elucidate the relationship between glycosylation of IgG and aging, oligosaccharide structures of human IgG purified from sera of men and women aged 18 to 73 years were investigated. Oligosaccharides were liberated quantitatively from IgG by hydrazinolysis followed by N-acetylation and were tagged with p-aminobenzoic acid ethyl ester. The oligosaccharide structures were then analyzed by HPLC in conjunction with sequential exoglycosidase digestion. All IgG samples were shown to contain a series of biantennary complex type oligosaccharides which consisted of ±Gal1-4GlcNAc1-2Man1-6(±GlcNAc1-4)(±Gal1-4GlcNAc1-2Man1-3)Man1-4GlcNAc1-4(±Fuc1-6)GlcNAc and their mono- and di-sialo glycoforms in different ratios. In female IgG samples only, the incidence of non-galactosylated oligosaccharides with non-reducing terminal GlcNAc residues increased with aging (r>0.8), whereas that of digalactosylated oligosaccharides decreased (r<–0.8). A weaker correlation was observed between aging and the incidence of neutral and monosialo oligosaccharides in female IgG (r:0.461 and r=–0.538, respectively) and between aging and the incidence of oligosaccharides with a bisecting GlcNAc in both male and female IgG samples (r=0.566 and r=0.440, respectively). In addition, a significant change with aging in the galactosylation of IgG oligosaccharides was observed in females in their thirties, fifties, and sixties (p<0.02, p<0.01, and p<0.04, respectively). These findings may contribute to our understanding of autoimmune diseases such as rheumatoid arthritis in which glycosylation is involved. 相似文献
20.
Ossi Renkonen Leena Penttilä Anne Makkonen Ritva Niemelä Anne Leppänen Jari Helin Anja Vainio 《Glycoconjugate journal》1989,6(1):129-140
A novel linear tetrasaccharide, Gal1-4GlcNAc1-6Gal1-4GlcNAc, was isolated from partial acid hydrolysates of metabolically labeled poly-N-acetyllactosaminoglycans of murine teratocarcinoma cells. It was characterized by exo-glycosidase sequencing and by mild acid hydrolysis followed by identification of all partial cleavage products. The tetrasaccharide, and likewise labelled GlcNAc1-6Gal1-4GlcNAc, resisted the action of endo--galactosidase (EC 3.2.1.103) fromE. freundii at a concentration of 125 mU/ml, while the isomeric, radioactive teratocarcinoma saccharides Gal1-4GlcNAc1-3Gal1-4GlcNAc and GlcNAc1-3Gal1-4GlcNAc were cleaved in the expected manner.Abbreviations WGA
wheat germ agglutinin
- BSA
bovine serum albumin
- [3H]GlcNAc1-4-GlcNAc1-4GlcNAcOL
N,N,NN'-triacetylchitotriose reduced with NaB3H4 相似文献