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1.
Boel Lanne Jeana Cîopraga Jörgen Bergström Cecilia Motas Karl-Anders Karlsson 《Glycoconjugate journal》1994,11(4):292-298
The carbohydrate-binding specificity ofPseudomonas aeruginosa lectin I (PA-I) in iodinated or biotinylated form was studied. A large number of glycosphingolipids, as well as some glycoproteins and neoglycoproteins were used as ligands. Also, inhibition by free saccharides of PA-I binding to glycosphingolipids was tested. It was found that the lectin binds most strongly to terminal and nonsubstituted Gal3Gal- or Gal4Gal-structures.Abbreviations PA-I
Pseudomonas aeruginosa lectin I
- Cer
ceramide
- lactosylceramide
Gal4GlcCer
- iso globotriaosylcerami
Gal3Gal4GlcCer
- globotriaosylceramide
Gal4Gal4GlcCer
- globoside or globotetraosylceramide
GalNAc3Gal4Gal4GlcCer
- Forssman glycolipid
GalNAc3GalNAc3Gal4Gal4GlcCer
- P1 glycolipid
Gal4Gal4GlcNAc3Gal4GlcCer
- lactoneotetraosylceramide
Gal4GlcNAc3Gal4GlcCer
- B5 glycolipid
Gal3Gal4GlcNAc3Gal4GlcCer
- gangliotetraosylceramide
Gal3GalNAc4Gal4GlcCer
- GM1
Gal3GalNAc4(NeuAc3)Gal4GlcCer
- RBC
red blood cells
- BSA
bovine serum albumin
- PBS
phosphate-buffered saline
- SDS
sodium dodecyl sulfate
- TLC
thin-layer chromatography
- HPLC
high pressure liquid chromatography
- MS
mass spectrometry
- FAB
fast-atom bombardment
- EI
electron impact 相似文献
2.
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
3.
Antti Seppo Leena Penttilä Anne Leppänen Hannu Maaheimo Ritva Niemelä Jari Helin Jean-Michel Wieruszeski Ossi Renkonen 《Glycoconjugate journal》1994,11(3):217-225
1,3-Galactosylation of radiolabelled bi-antennary acceptors Gal1-4GlcNAc1-3(Gal1-4GlcNAc1-6)Gal-R (R=1-OH, 1-4GlcNAc or 1-4Glc) with bovine thymus 1,3-galactosyltransferase was studied. At all stages of the reactions the three acceptors reacted faster at the 1 6 linked arm than at the 1 3 linked branch. Hence, in addition to the doubly 1,3-galactosylated products, practically pure Gal1-4GlcNAc1-3(Gal1-3Gal1-4GlcNAc1-6)Gal-R could be obtained from the three acceptors in reactions that had proceeded to near completion. The isomeric mono-1,3-galactosylated products were identified by using exoglycosidases to remove the branches unprotected by 1,3-galactoses and by subsequently identifying the resulting linear glycans chromatographically.Abbreviations Gal
d-galactose
- GlcNAc
N-acetyl-d-glucosamine
- Lac
lactose
- LacNAc
Gal1-4GlcNAc
- MH
maltoheptaose
- MP
maltopentaose
- MT
maltotriose
- MTet
maltotetraose
- WGA
wheat germ agglutinin
- 3
position 3 of the galactose unit of LacNAc or Lac
- 6
position 6 of the galactose unit of LacNAc or Lac 相似文献
4.
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
5.
Summary A mouse embryonic stem (ES) cell line E14 and early mouse embryos were stained with a panel of 15 monoclonal antibodies recognizing sialylated or potentially sialylated carbohydrate determinants, Sialyl Le-x and sialyl Le-a were detected on the pre-implantation embryo from the 8-cell stage, and sialyl Le-a weakly on undifferentiated ES cells. Changes in cell surface carbohydrates occurred after induction of ES cell differentiation with retinoic acid (RA) and dibutyryl cAMP. Qualitative analysis of the neutral glycolipids of untreated and RA-treated ES cells using high-performance thin-layer chromatography (HPTLC) revealed few differences between the two types of culture. The major gangliosides in both cultures were indicative of an active a ganglioside synthesis pathway. GD3, a precursor of the b synthesis pathway, previously reported to be characteristic of embryonal carcinoma (EC) cells, was absent. RA-induced differentiation caused a shift in the spectrum to more complex gangliosides. Application of fast atom bombardment mass spectrometry (FAB-MS) to permethylated derivatives of individual bands permitted partial characterization of an unusual sialylated glycolipid and a rare ganglioside with the suggested structure of GalNAc-GD1a.Abbreviations NeuAc
N-acetylneuraminic acid
- Cer
ceramide
- CMH
monohexosylceramide
- CDH
lactosylceramide (Gal1-4Glc1-Cer)
- CTH
ceramide trihexoside (Gal1-4Gal1-4Glc1-Cer)
- globoside
(GalNAc1-3 Gal1-4Gal1-4Glc1-Cer)
- Forssman antigen
(GalNAc1-3GalNAc1-3Gal1-4Gal1-4Glc1-Cer)
- GM3
(NeuAc2-3Gal1-4Glc1-Cer)
- GD3
(NeuAc2-8NeuAc2-3Gal1-4Glc1-Cer)
- GM1
(Gal1-3GalNAc1-4[NeuAca2-3]Gal1-4Glc1-Cer)
- GD1a
(NeuAc2-3Gal1-3GalNAc1-4[NeuAc2-3]Gal1-4Glc1-Cer)
- GT1b
(Neu5Ac2-3Gal1-3GalNAc1-4[Neu5Ac2-8Neu5Ac2-3]Gal1-4Glc1-Cer)
The glycolipids are named according to Svennerholm (1963) and the recommendations of the IUPAC-IUB Commission on Biochemical Nomenclature (1978). 相似文献
6.
Guiyun Xu Takashi Suzuki Yasuhiro Maejima Tohru Mizoguchi Mamoru Tsuchiya Makoto Kiso Akira Hasegawa Yasuo Suzuki 《Glycoconjugate journal》1995,12(2):156-161
The sialidase of swine influenza A viruses of N1 and N2 subtypes, isolated from 1930 to 1992, was studied for substrate specificity with ganglio-series, lacto-series type II and GM3 gangliosides containing Neu5Ac2-3Gal, Neu5Gc2-3Gal and Neu5Ac2-6Gal linkages. All viral sialidases tested showed that the activity for hydrolysing substrates with Neu5Ac2-3Gal was higher than the activities with Neu5Gc2-3Gal and Neu5Ac2-6Gal linkages. When GM1b, GM3 and sialylparagloboside were used as substrates, the earliest strain (A/Wisconsin/15/30 H1N1, isolated in 1930) showed the activity ratio of Neu5Ac2-6Gal to Neu5Ac2-3Gal to be 0.13:0.2, and the ratio Neu5Gc2-3Gal/Neu5Ac2-3Gal to be 0.19:0.37, while those strains isolated from 1978 to 1992 exhibited ratios of 0.29:0.58 for Neu5Ac2-6Gal/Neu5Ac2-3Gal and 0.51:0.76 for Neu5Gc2-3Gal/Neu5Ac2-3Gal. The above results indicate that the substrate specificities of sialidases from swine influenza A viruses towards sialyl linkages and the molecular species of sialic acid are related to the year of isolation, i.e. strains isolated after 1978 exhibited higher activity towards Neu5Ac2-6Gal and Neu5Gc2-3Gal linkages when compared with strains isolated in an earlier year, 1930.Abbreviation Neu5Ac
5-N-acetylneuraminic acid
- Neu5Gc
5-N-glycolyneuraminic acid
- Gal
d-galactose
- Glc
d-glucose
- Cer
Ceramide
- II3(Neu5Ac)Lac
Neu5Ac2-3Gal1-4Glc
- GM3(Neu5Ac2-3Gal)
Neu5Ac2-3Gal1-4Glc1-Cer
- GM3(Neu5Gc2-3Gal)
Neu5Gc2-3Gal1-4Glc1-Cer
- GM1b(Neu5Ac2-3Gal)
Neu5Ac2-3Gal1-3GalNac1-4Gal1-4Glc1-Cer
- GMlb(Neu5Gc2-3Gal)
Neu5Gc2-3Gal1-3GalNAc1-4Gal1-4Glc1-Cer
- IV3(Neu5Ac)nLc4Cer
Neu5Ac2-3Gal1-3GlcNAc1-4Gal1-4Glc1-Cer
- IV3(Neu5Gc)nLc4Cer
Neu5Gc2-3Gal1-3GlcNAc1-4Gal1-4Glc1-Cer
- IV6(Neu5Ac)nLc4Cer
Neu5Ac2-6Gal1-3GlcNAc1-4Gal1-4Glc1-Cer
- TDC
taurodeoxycholate. 相似文献
7.
Enzymatic characterization of CMP-NeuAc:Galβ1-4GlcNAc-R α(2-3)-sialyltransferase from human placenta
In this report we present the enzymatic characterization of CMP-NeuAc:Gal1-4GlcNAc-R (2-3)-sialyltransferase from human placenta using placenta membranes as an enzyme preparation. This sialyltransferase is highly sensitive to detergents and prefers type 2 chain (Gal1-4GlcNAc) over type 1 chain (Gal1-3GlcNAc) acceptors. Oligosaccharides and glycopeptides were better acceptor substrates than glycoproteins. Of the branched oligosaccharides, those with a bisectedN-acetylglucosamine (GlcNAc) structure appeared to be poorer substrates, while triantennary structures containing a Gal1-4GlcNAc1-4Man1-3Man branch were preferred. Product characterization, using 400 MHz1H-NMR spectroscopy, confirmed that sialic acid was introduced into the Gal1-4GlcNAc-R units of the acceptor substrates in an (2-3) linkage, and revealed that this sialytransferase does not prefer either of the two branches of a complex type diantennary glycopeptide acceptor for sialic acid attachment. These properties distinguish this enzyme from all other sialyltransferases characterized to date.Abbreviations NeuAc
N-acetylneuraminic acid
- CMP-NeuAc
cytidine 5-monophospho-N-acetylneuraminic acid
- GP-F2 and GP-F4
diantennary complex type glycopeptides from desialylated fibrinogen
- GP-Trf
diantennary complex type glycopeptide from desialylated transferrin
- LNT
Gal1-3GlcNAc1-3Gal1-4Glc (lacto-N-tetraose)
- 6-sialytransferase
CMP-NeuAc:Gal1-4GlcNAc-R (2-6)-sialytransferase
- 3-sialytransferaseO
CMP-NeuAc:Gal1-3GalNAc-R (2-3)-sialyltransferase
- 3-sialytransferase I
CMP-NeuAc:Gal1-3(4)GlcNAc-R (2-3)-sialyltransferase
- 3-sialytransferase II
CMP-NeuAc:Gal1-4GlcNAc-R (2-3)-sialytransferase 相似文献
8.
Marti F A Bierhuizen Hommo Tedzes Wietske E C M Schiphorst Dirk H van den Eijnden Willem van Dijk 《Glycoconjugate journal》1988,5(1):85-97
The effects of branching and substitution of branches by sialic acid and fucose on the interaction ofN-linked glycopeptides and related oligosaccharides with immobilizedPhaseolus vulgaris leukoagglutinating lectin (L-PHA) were examined. Asialo bi-, tri-and tetra-antennary glycans were all retarded but to different extents on a long column of L-PHA-agarose. Asialo tri- and tetra-antennary glycans containing the pentasaccharide unit Gal1-4GlcNAc1-2[Gal1-4GlcNAc1-6]Man were strongly retarded, whereas asialo bi- and tri-antennary glycans lacking the Gal1-4GlcNAc1-6 branch were only weakly retarded. In all instances the interaction with the lectin was completely abolished when either (2–6)-linkedN-acetylneuraminic acid or (1–3)-linked fucose was present at the galactose orN-acetylglucosamine residue of the Gal1-4GlcNAc1-6Man1-6 branch, respectively. The same substitutions on the Gal1-4GlcNAc1-6Man1-6 branch decreased but did not abolish the affinity of the lectin for the glycans. The presence of NeuAc2-6 and Fuc1-3 on the other two branches did not interfere with the binding of the glycans to L-PHA. Furthermore, it appeared that the presence of the Man1-4GlcNAc unit is requried for interaction with the lectin. In order to obtain reliable information on the relative occurrence of tri- and tetra-antennary glycopeptides, this study shows that it is essential to desialylate and to defucosylate the glycans prior to application to L-PHA-agarose.Abbreviations L-PHA
leukoagglutinating phytohemagglutinin
- CMP-NeuAc
cytidine-5-monophospho-N-acetylneuraminic acid
-
GP
glycopeptide
-
OS
oligosaccharide
- HPLC
high-performance liquid chromatography
- FNR
fraction not retarded
- FR
fraction retarded
suffixes MS, BS and TS indicate mono-, bi- and trisialyl derivatives respectively; suffix MF indicates monofucosyl derivatives.structures of the substratesOS2, OS3, OS3, OS4, GP2, GP3, GP4, GP4-MF, OS2(3) andOS2(-) are presented in Fig. 2 相似文献
9.
We report here the in vivo production of type 2 fucosylated-N-acetyllactosamine oligosaccharides in Escherichia coli. Lacto-N-neofucopentaose Gal1-4GlcNAc1-3Gal1-4(Fuc1-3)Glc, lacto-N-neodifucohexaose Gal1-4(Fuc1-3)Glc-NAc1-3Gal1-4(Fuc1-3)Glc, and lacto-N-neodifucooctaose Gal1-4GlcNAc1-3Gal1-4(Fuc1-3)GlcNAc1-3Gal1-4(Fuc1-3)Glc were produced from lactose added in the culture medium. Two of them carry the Lewis X human antigen. High cell density cultivation allowed obtaining several grams of fucosylated oligosaccharides per liter of culture. The fucosylation reaction was catalyzed by an -1,3 fucosyltransferase of Helicobacter pylori overexpressed in E. coli with the genes lgtAB of N. meningitidis. The strain was genetically engineered in order to provide GDP-fucose to the system, by genomic inactivation of gene wcaJ involved in colanic acid synthesis and overexpression of RcsA, positive regulator of the colanic acid operon.To prevent fucosylation at the glucosyl residue, lactulose Gal1-4Fru was assayed in replacement of lactose. Lactulose-derived oligosaccharides carrying fucose were synthesized and characterized. Fucosylation of the fructosyl residue was observed, indicating a poor acceptor specificity of the fucosyltransferase of H. pylori. 相似文献
10.
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
11.
Mark A. Williams Hiroshi Kitagawa Arun K. Datta James C. Paulson James C. Jamieson 《Glycoconjugate journal》1995,12(6):755-761
Values ofK
m were determined for three purified sialyltransferases and the corresponding recombinant enzymes. The enzymes were Gal1-4GlcNAc 2-6sialyltransferase and Gal1-3(4)GlcNAc 2-3sialyltransferase from rat liver; these enzymes are responsible for the attachment of sialic acid to N-linked oligosaccharide chains; and the Gal1-3GalNAc 2-3sialyltransferase from porcine submaxillary gland that is responsible for the attachment of sialic acid to O-linked glycoproteins and glycolipids. A procedure for the large scale expression of active sialyltransferases from recombinant baculovirus-infected insect cells is described. For the liver enzymes values ofK
m were determined using rat and human asialo1 acid glycoprotein andN-acetyllactosamine as variable substrates; lacto-N-tetraose was also used with the Gal1-3(4)GlcNAc 2-3sialyltransferase. Antifreeze glycorprotein was used as the macromolecular acceptor for the porcine enzyme. Values forK
m were also determined using CMP-NeuAc as the variable substrate.Abbreviations NeuAc
N-acetylneuraminic acid
- Gal
galactose
- GlcNAc
N-acetylglucosamine
Enzymes: Gal1-4GlcNAc 2-6sialyltransferase, EC 2.4.99.1; Gal1-3(4)GlcNAc 2-3sialyltransferase, EC 2.4.99.5; Gal1-3GalNAc 2-3sialyltransferase, EC 2.4.99.4. 相似文献
12.
Sarah Ehrlich-Rogozinski Antonio De Maio Halina Lis Nathan Sharon 《Glycoconjugate journal》1987,4(4):379-390
The interaction of125I-labeledErythrina cristagalli agglutinin (ECA) with neutral glycosphingolipids on thin layer chromatograms was examined by the overlay technique followed by radioautography. The lectin bound topara-globoside with a sensitivity about 10 times higher than to lactosylceramide or globoside, in agreement with the specificity of the lectin forN-acetyllactosamine. The lower limit of detection ofpara-globoside was about 0.66 nmol. The specific binding of ECA to this glycolipid was confirmed by a highly sensitive enzyme-linked lectin assay (ELLA), utilizing the horseradish peroxidase-avidin-biotin system for detection of bound lectin. Overlays of neutral glycosphingolipid extracts from human erythrocyte membranes and from human granulocytes with ECA demonstrated that the lectin can be employed for the detection of small amounts ofpara-globoside in biological materials also in the presence of excess globoside. No staining was obtained when thin layer chromatograms of neutral glycosphingolipid extracts from rabbit erythrocyte membranes were overlayed with125I-ECA. Afterin situ treatment of the chromatograms with -galactosidase, the lectin bound to several components, one of which had a mobility corresponding to that of the pentahexosylceramide Gal3Gal4GlcNAc3Gal4Glc1Cer, the major neutral glycosphingolipid of rabbit erythrocytes, thus providing further evidence for the specificity of ECA forpara-globoside.Abbreviations GSL
glycosphingolipid(s)
- CDH
lactosylceramide, Gal4Glc1Cer
- CTH
trihexosylceramide, Gal4Gal4Glc1Cer
- GLOB
globoside, GalNac3Gal4Gal4Glc1Cer
- PG
para-globoside, Gal4GlcNAc3Gal4Glc1Cer
- AsGM1
asialo-GM1, Gal3GalNAc4Gal4Glc1Cer
- FORS
Forsmann antigen, GalNAc3GalNAc3Gal4Gal4Glc1Cer
- CPH
pentahexosylceramide, Gal3Gal4GlcNAc3Gal4Glc1Cer
- ECA
Erythrina cristagalli agglutinin
- SBA
soybean agglutinin
- PBS
phosphate-buffered saline
- PVP-40
polyvinylpyrrolidone M.W. 40000
- BSA
bovine serum albumin
- HRP-avidin
horseradish peroxidase conjugated to avidin
- ELLA
enzyme-linked lectin assay
- ELISA
enzyme-linked immunosorbent assay
- PMNL
polymorphonuclear leukocytes
- HPTLC
high performance thin layer chromatography 相似文献
13.
The regional difference in the carbohydrate components of the ductus epididymis epithelium of a lizard was delineated by means of 13 lectins. Basal cells expressed only N-acetylglucosamine (GlcNAc). Throughout the ductus, the secretory cells showed oligosaccharides with terminal N-acetylneuraminic acid (Neu5Ac)(2,6)galactose (Gal)/N-acetylgalactosamine (GalNAc) and internal mannose (Man) and/or glucose (Glc) in the whole cytoplasm, oligosaccharides terminating in Neu5Ac(2,6)Gal(1,3)GalNAc, Neu5Ac(2,6)Gal (1,4)GlcNAc, GalNAc, GlcNAc, and fucose (Fuc) in the supra-nuclear zone, and also glycans terminating in Neu5Ac(2,3)Gal (1,4)GlcNAc, Neu5Ac(2,6)Gal(1,3)GalNAc, Gal (1,4)GlcNAc on the luminal surface. In the caput and corpus regions, the supra-nuclear cytoplasm was characterized by terminal Gal(1,4)GlcNAc and GalNAc, the luminal surface by GalNAc and Gal. The Golgi zone, showing oligosaccharides with terminal Neu5Ac(2,3)Gal (1,4)GlcNAc, Neu5Ac(2,6)Gal (1,3)GalNAc, Neu5Ac(2,6)Gal (1,4)GlcNAc, and internal GlcNAc, expressed terminal Gal (1,4)GlcNAc and GalNAc in the caput, and terminal GalNAc in the corpus. The granules showed all the investigated carbohydrates in their peripheral zone except terminal GalNAc and Fuc, whereas internal Man/Glc and terminal Gal were expressed in the central core, and Fuc throughout the ductus, terminal GlcNAc in the caput and corpus, and terminal GalNAc only in the corpus. 相似文献
14.
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
15.
-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
16.
Molecular dynamics simulations of models of unmodified and deiminated MBP (myelin basic protein) have been performed on solvated structures with added counterions, for 10 ns using AMBER (assisted model building with energy refinement). The protein structures became extended, and a considerable number of -helical segments formed spontaneously. The degree of molecular extension was greater in the deiminated species, and the -helices were more transient. These structural disruptions may be operative in vivo during multiple sclerosis.Figure A model of the C1 isoform of myelin basic protein showing major -helical segments that were stable over the last 1 ns of a molecular dynamics simulation. The -helices are colored red, the -strands are colored yellow, the -turns are colored blue, and random coils are colored green 相似文献
17.
Philippe Delannoy Isabelle Kim Nathalie Emery Carmen de Bolos Andre Verbert Pierre Degand Guillemette Huet 《Glycoconjugate journal》1996,13(5):717-726
We have analysed the mucins synthesized by the HT-29 MTX cell subpopulation, derived from the HT-29 human colon carcinoma cells through a selective pressure with methotrexate (Lesuffleuret al., 1990,Cancer Res
50: 6334–43), in the presence of benzyl-N-acetyl--galactosaminide (GalNAc-O-benzyl), which is a potential competitive inhibitor of the 1,3-galactosyltransferase that synthesizes the T-antigen. The main observation was a 13-fold decrease in the sialic acid content of mucins after 24 h of exposure to 5mm GalNAc-O-benzyl. This effect was accompanied by an increased reactivity of these mucins to peanut lectin, testifying to the higher amount of T-antigen. The second observation was a decrease in the secretion of the mucins by GalNAc-O-benzyl treated cells. The decrease in mucin sialyation was achieved through thein situ -galactosylation of GalNAc-O-benzyl into Gal1–3GalNAc-O-benzyl, which acts as a competitive substrate of Gal1–3GalNAc 2,3-sialyltransferase, as shown by the intracellular accumulation of NeuAc2–3Gal1–3GalNAc-O-benzyl in treated cells.Abbreviations BSM
bovine submaxillary mucin
- MTX
methotrexate
- PBS
sodium phosphate 10mm, NaCl 0.15m, pH 7.4 buffer
-
pNp
p-nitrophenol
- TBS
Tris/HCl 10mm, NaCl 0.15m, pH 7.4 buffer
Enzymes: CMP-NeuAc: Gal1–3/4GlcNAc 2,3-sialyltransferase, ST3(N), EC 2.4.99.6; CMP-NeuAc: Gal1–4GlcNAc 2,6-sialyltransferase, ST6(N), EC 2.4.99.1; CMP-NeuAc: Gal1–3GalNAc 2,3-sialyltransferase, ST3(O), EC 2.4.99.4; CMP-NeuAc: R-GalNAc1-O-Ser 2,6-sialyltransferase, ST6(O)-I, EC 2.4.99.3; CMP-NeuAc: NeuAc2–3Gal1–3GalNAc 2,6-sialyltransferase, ST6(O)-II, EC 2.4.99.7; UDP-GlcNAc: Gal1–3GalNAc-R·(GlcNAc to GalNAc) 1,6-N-acetylglucosaminyltransferase, EC 2.4.1.102; UDP-GlcNAc: GalNAc-R 1,3-N-acetylglucosaminyltransferase, EC 2.4.1.147; UDP-Gal: GalNAc-R 1,3-galactosyltransferase, EC 2.4.1.122. 相似文献
18.
The effect of cerulenin on the production of -lactamase and other periplasmic proteins was studied in Escherichia coli IA199 carrying plasmid pBR322. Cerulenin (10 to 25 g/ml) had almost no effect on the growth rate of E. coli but it decreased the amount of -lactamase and other periplamic proteins in shock fluid. Higher amounts of the antibiotic (40 to 100 g/ml)decreased turbidity and almost completely prevented synthesis of -lactamase and other periplasmic proteins. Cerulenin decreased incorporation of l-[35S]methionine into membranes during growth as well. Spheroplasts secreted -lactamase into the external medium, but during a 3-h incubation in the presence of cerulenin (25 g/ml) this secretion was prevented by more than 90%. -Lactamase was secreted into the isolated membrane vesicles from E. coli IA199. However, only 5% of the total amount of pre--lactamase was secreted and processed by the membranes in vitro. Cerulenin did not prevent processing in vitro but the membranes prepared from the cells grown in the presence of cerulenin (25 g/ml) did not catalyze processing of pre--lactamase at all. Membrane preparations from Bacillus subtilis did not process pre--lactamase either in the absence or in the presence of cerulenin. 相似文献
19.
Sreedhara Sangadala Subramanian Sivakami Joseph Mendicino 《Molecular and cellular biochemistry》1991,101(2):125-143
Summary Two specific -N-acetylglucosaminyltransferases involved in the branching and elongation of mucin oligosaccharide chains, namely, a 1,6 N-acetylglucosaminylsaminyltransferase that transfers N-acetylglucosamine from UDP-N-acetylglucosamine to Gal3GalNAc-Mucin to yield Gal3(GlcNAc6)GalNAc-Mucin and a 3-N-acetylglucosaminyl transferase that transfers N-acetylglucosamine from UDP-N-acetylglucosamine to Gal3(GlcNAC6)GalNAc-mucin to yield GlcNAc3Gal3 (GlcNAc6)GalNAc-Mucin were purified from the microsomal fraction of swine trachea epithelium. The 1,6-N-acetylglucosaminyltransferase was purified about 21,800-fold by procedures which included affinity chromatography on DEAE columns containing bound asialo Cowper's gland mucin glycoprotein with Gal1,3GalNAc side chains. The apparent molecular weight estimated by gel filtration was found to be about 60 Kd. The purified enzyme showed a high specificity for Gal1,3GalNAc chains and the most active substrates were mucin glycoproteins containing these chains. The apparent Km of the 6-glucosaminyltrans-ferase for Cowper's gland mucin glycoprotein containing Gal1,3GalNAc chains was 0.53 µM; for UDP-N-acetylglucosamine, 12 µM; and for Gal 1,3GalNAc NO2ø, 4 mM. The activity of the 6-glucosaminyltransferase was dependent on the extent of glycosylation of the Gal3GalNAc chains in Cowper's gland mucin glycoprotein.The best substrate for the partially purified 3-Glucosaminyltransferase was Cowper's gland mucin glycoprotein containing Gal1,3(GlcNAc6)GalNAc side chains. This enzyme showed little or no activity with intact sialylated Cowper's gland mucin glycoprotein or derivatives of this glycoprotein containing GalNAc or Gal1,3GalNAc side chains.The radioactive oligosaccharides formed by these enzymes in large scale reaction mixtures were released from the mucin glycoproteins by treatment with alkaline borohydride, isolated by gel filtration on Bio-Gel P-6 and characterized by methylation analysis and sequential digestion with exoglycosidases. The oligosaccharide products formed by the 6- and 3-glucosaminyltransferases were shown to be Gal3(GlcNAC6) GalNAc and GlcNAc3 Gal3(GlcNAC6)GalNAc respectively.Taken collectively, these results demonstrate that swine trachea epithelium contains two specific N-acetylglucosaminyltransferases which catalyze the initial branching and elongation reactions involved in the synthesis of O-linked oligosaccharide chains in respiratory mucin glycoproteins. The first enzyme a 6-glucosaminyltransferase converts Gal3GalNAc chains in mucin glycoproteins to Gal3(GlcNAc6)GalNAc chains. This product is the substrate for a second 3-glucosaminyltransferase which converts the Gal3(GlcNAc6)GalNAc chains to GlcNAc3Gal(GlcNAc6)GalNAc chains in the glycoprotein. The 3-glucosaminyltransferase did not utilize Gal3GalNAc chains as a substrate and this results in an ordered sequence of addition of N-acetylglucosamine residues to growing oligosaccharide chains in tracheal mucin glycoproteins.Abbreviations NeuNAc
N-acetylneuraminic acid
- GalNAcol
N-acetylgalactosaminitol
- CGMG
Cowper's gland mucin glycoprotein
- GalNAc-CGMG
Cowper's gland mucin glycoprotein containing GalNAc side chains O-glycosidically linked to serine or threonine
- Gal3GalNAc-CGMC
Cowper's gland mucin glycoprotein containing Gal3GalNAc side chains
- MES
2-(N-morpholino) Ethane Sulfonic acid
- PBS
Phosphate Buffered Saline 相似文献
20.
Albert M. Wu 《Molecular and cellular biochemistry》1984,61(2):131-1411
Summary The binding properties of Arachis hypogaea (PNA), Bauhinia vurpurea alba (BPL), Maclura pomifera (MPL) and Sophora japonica (SJL) lectins were studied by quantitative precipitin and precipitin inhibition assays, demonstrating them to be most specific for dGal13dGalNAc residues. Additionally, each lectin had its own binding characteristic such as different binding abilities to dGal14dGlcNAc or dGal13dGlcNAc1linked oligosaccharides, and/or dGalNAc1linked to the Ser or Thr of the protein moiety. These differential binding characteristics can be used for investigating fine differences of the carbohydrate structure of the glycoconjugates, especially those having dGal13dGalNAc residues as terminal non-reducing ends.Abbreviations
dGal
d-galactopyranose
-
dMan
d-mannopyranose
-
dGalNAc
2-acetamido-2-deoxy-d-galacto-pyranose
-
dGlcNAc
2-acetamino-2-deoxy-d-glucopyranose
- LFuc
L-fucose
- NeuNAc
N-acetylneuraminic acid
- Ser
serine
- Thr
Threonine
- RCA
Ricinus communis agglutinin
- SBA
Soy bean agglutinin (Glycine max)
- HPA
Helix pomatia agglutinin
- DBL
Dolichos biflorus lectin
- GCL
Geodia cydonium lectin 相似文献