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1.
The125I-labeled fragment C of tetanus toxin was found to bind specifically to the gangliosides GD1b, GT1b, and GQ1b when applied to thin-layer chromatograms on which a mixture of gangliosides had been resolved. As little as 2.5 pmoles of these gangliosides could be detected by this method. In addition to factors determined by the sample, namely the amount and species of gangliosides present, optimal binding of the125I-labeled fragment C also depended upon the iodination procedure used to generate the probe, the toxin concentration, and the concentration, buffer type, pH, and ionic strength of the binding solution. This new technique was shown to be a sensitive method for the detection and identification of specific gangliosides originating from extraneural or neural cells.Nomenclature: The gangliosides follow the nomenclature system of Svennerholm [Eur J Biochem (1977) 79:11–21] GM3
II3NeuAc-LacCer
- GD3
II3(NeuAc)2-LacCer
- GM1
II3NeuAc-GgOse4Cer
- GD1a
IV3NeuAc, II3NeuAc-GgOse4Cer
- GD1b
II3(NeuAc)2-GgOse4Cer
- GT1b
IV3NeuAc, II3(NeuAc)2-GgOse4Cer
- GQ1b
IV3(Neu-Ac)2, II3(NeuAc)2-GgOse4Cer
- GP1b
IV3(NeuAc)3, II3(NeuAc)2-GgOse4Cer 相似文献
2.
Olle Nilsson Jan-Erik MÅnsson Tomas Brezicka Jan Holmgren Leif Lindholm Sverre Sörenson Fridrik Yngvason Lars Svennerholm 《Glycoconjugate journal》1984,1(1):43-49
Characterization of monosialogangliosides of a small cell lung carcinoma showed a unique composition. The tumour contained GM2 and Fucosyl-GM1 (Fuc-GM1) with 2-hydroxy fatty acids as major ganglioside components. Three out of four other small cell carcinomas analysed contained also Fuc-GM1 as a characteristic ganglioside. Fuc-GM1 is suggested to be a small-cell lung carcinoma associated ganglioside antigen.Nomenclature: The gangliosides have been designated according to Svennerholm [25] GM3
II3NeuAc-Lac-Cer
- GD3
II3(NeuAc)2-LacCer
- GM2
II3NeuAc-GgOse3Cer
- GM1
II3NeuAc-GgOse4Cer
- Fuc-GM1
FuclV2Neu-AcII3-GgOse4Cer
- 3-LM1
IV3NeuAc-nLcOse4Cer
- 6-LM1
IV6NeuAc-nLcOse4Cer 相似文献
3.
Johannes Müthing Jasna Peter-Katalinić Franz-Georg Hanisch Frank Unland Jürgen Lehmann 《Glycoconjugate journal》1994,11(2):153-162
The aim of the present study was to investigate the ganglioside expression of the highly metastatic murine lymphoreticular tumour cell line MDAY-D2. Cells were propagated under controlled pH conditions and oxygen supply in bioreactors of 1 and 7.5l volumes by repeated batch fermentation. Gangliosides were isolated from 2.7×1011 cells, purified by silica gel chromatography and separated into mono- and disialoganglioside fractions by preparative DEAE anion exchange high performance liquid chromatography. Individual gangliosides were obtained by preparative thin layer chromatography. Their structural features were established by immunostaining, fast atom bombardment and gas chromatography mass spectrometry. In addition to gangliosides of the GM1a-pathway (GM2, GM1a and GD1a) and GM1b (IV3Neu5Ac-GgOse4Cer) and GalNAc-GM1b of the GM1b-pathway, the dis8aloganglioside GD1 (IV3Neu5Ac, III6Neu5Ac-GgOse4Cer) was found in equal amounts compared to GD1a (IV3Neu5Ac, II3Neu5Ac-GgOse4Cer). All gangliosides were substituted with C24:0,24:1 and C16:0 fatty acids, sphingosine andN-acetylneuraminic acid as the sole sialic acid.
Abbreviations: FAB-MS, fast atom bombardment-mass spectrometry; GC-MS, gas chromatography-mass spectrometry; GSL(s), glycosphingolipid(s); HPLC, high performance liquid chromatography; HPTLC, high performance thin layer chromatography; Neu5Ac,N-acetylneuraminic acid; Neu5Gc,N-glycoloylneuraminic acid [57]. The designation of the following glycosphingolipids follows the IUPAC-IUB recommendations [58] and the nomenclature of Svennerholm [59]. Gangliotriaosylceramide or GgOse3Cer, GalNAc1-4Gal1-4Glc1-1Cer; gangliotetraosylceramide or GgOse4Cer, Gal1-3GalNAc1-4Gal1-4Glc1-1Cer gangliopentaosylceramide or GgOse5Cer, GalNAc1-4Gal1-3GalNAc1-4Gal1-4Glc1-1Cer; GM2, II3Neu5Ac-GgOse3Cer; GM1a, II3Neu5Ac-GgOse4Cer; GM1b, IV3Neu5Ac-GgOse4Cer; GalNAc-GM1b, IV3Neu5Ac-GgOse5Cer; GD1a, IV3Neu5Ac, II3Neu5Ac-GgOse4Cer; GD1b, II3(Neu5Ac)2-GgOse4Cer; GD1 or GD1e, IV3Neu5Ac, III6Neu5AcGgOse4Cer; GD1e, IV3(Neu5Ac)2-GgOse4Cer; GT1b, IV3Neu5Ac, II3(Neu5Ac)2-GgOse4Cer. 相似文献
4.
Non-acid and acid glycosphingolipids were isolated from feces of one litter of germ-free rats from day 17 to day 51. Quantitative and qualitative changes described for small intestine of conventional rats [Bouhours D, Bouhours J-F (1981) Biochem Biophys Res Commun 99:1384–89] were also found in the feces of these germ-free rats. A decrease in lactosylceramide and sialyllactosylceramide excretion and a change fromN-acetylneuraminic acid toN-glycoloylneuraminic acid, as well as an appearance of type 1 chain blood group H-active penta- and decaglycosylceramides were observed during the weaning period. Thus the dramatic changes seen in rat intestinal glycosphingolipids postnatally seem to be primarily regulated by non-microbial factors.Abbreviations GM3
GM3-ganglioside, II3NeuAc-LacCer or II3NeuGc-LacCer
- SPG
IV3NeuAc-nLcOse4Cer
- GM1
GM1-ganglioside, II3NeuAc-GgOse4Cer 相似文献
5.
Johannes Müthing Jasna Peter-Katalinić Franz-Georg Hanisch Ulrich Neumann 《Glycoconjugate journal》1991,8(5):414-423
YAC-1 cells were propagated in bioreactors in 11 and 7.51 volumes. The cells were metabolically labelled withd-[1-14C]galactose andd-[1-14C]glucosamine. The ganglioside fraction, purified by DEAE-Sepharose and silica gel column chromatography, showed on thin layer chromatography four major bands with mobilities between GM1 and GD1a. Gangliosides, obtained by further purification steps including high performance liquid chromatography on silica gel 60 columns with a gradient system of isopropanol:hexane:water, and preparative high performance TLC were characterized by (1) immunostaining of corresponding asialogangliosides obtained by mild acid hydrolysis and neuraminidase treatment and (2) fast atom bombardment mass spectrometry of native and permethylated samples and methylation analysis of GM1b ganglioside. As well as small amounts of GM2 and GM1, the major gangliosides found in the complex mixture were GM1b and GalNAc-GM1b. The structural heterogeneity of these gangliosides was cased by (a) substitution of the ceramide moiety by fatty acids of different chain length and degree of unsaturation (C16:0, C24:0, C24:1) and (b) N-substitution of the sialic acid moieties with either acetyl or glycolyl groups. Disialogangliosides were detected only in low amounts and will be the subject of further investigation. A polyclonal chicken antiserum was raised against IVNeuAc-GgOse5Cer. The antiserum was highly specific for gangliosides (IVNeuAc and IVNeuGc) and asialogangliosides with a GgOse5Cer backbone. No cross-reaction with GM1b or GgOse4Cer was observed.
Abbreviations: FAB-MS, fast atom bombardment mass spectrometry; GSL(s), glycosphingolipid(s); HPLC, high performance liquid chromatography, HPTLC, high performance thin layer chromatography; NK, natural killer; SIM, selective ion monitoring; TIC, total ion current. NeuAc,N-acetylneuraminic acid; NeuGc,N-glycolylneuraminic acid. The designation of the following glycosphingolipids follows the IUB-IUPAC recommendations. GgOse3Cer or gangliotriaosylceramide or asialo GM2, GalNAc1-4Gal1-4GlcCer; GgOse4Cer or gangliotetraosylceramide or asialo GM1, Gal1-3GalNAc1-4Gal1-4GlcCer; GgOse5Cer organgliopentaosylceramide, GalNAc1-4Gal1-3GalNAc1-4Gal1-4GlcCer; II3NeuAc-GgOse4Cer or GM1; IV3NeuAcGgOse4Cer or GM1b; IV3NeuAc-GgOse5Cer or GalNAc-GM1b; IV3NeuAc, II3NeuAc-GgOse4Cer or GD1a; II3(NeuAc)2-GgOse4Cer or GD1b; IV3(NeuAc)2-GgOse4Cer or GD1c; IV3NeuAc,III6NeuAc-GgOse4Cer or GD1a; IV3NeuAc,II3(NeuAc)2-GgOse4Cer or GT1b;Vibrio cholerae and Arthrobacter ureafaciens neuraminidase (EC 3.2.1.18). 相似文献
6.
Johannes Müthing Frank Unland Dagmar Heitmann Michaela Orlich Franz-Georg Hanisch Jasna Peter-Katalinić Vera Knäuper Harald Tschesche Sørge Kelm Roland Schauer Jürgen Lehmann 《Glycoconjugate journal》1993,10(1):120-126
The structures of gangliosides from human granulocytes were elucidated by fast atom bombardment mass spectrometry and by gas chromatography/mass spectrometry as their partially methylated alditol acetates. In human granulocytes besides GM3 (II3Neu5Ac-LacCer), neolacto-series gangliosides (IV3Neu5Ac-nLcOse4Cer, IV6Neu5Ac-nLcOse4Cer and VI3Neu5Ac-nLcOse6Cer) containing C24:1, and to some extent C22:0; and C16:0 fatty acid in their respective ceramide portions, were identified as major components. In this study we demonstrate that gangliosides from human granulocytes, the second most abundant cells in peripheral blood, can serve as receptors for influenza viruses A/PR/8/34 (H1N1), A/X-31 (H3N2), and a parainfluenza virus Sendai virus (HNF1, Z-strain). Viruses were found to exhibit specific adhesion to terminal Neu5Ac2-3Gal and/or Neu5Ac2-6Gal sequences as well as depending on the chain length of ganglioside carbohydrate backbones from human granulocytes, these important effector cells which represent the first line of defence in immunologically mediated reactions.
Abbreviations: FAB-MS, fast atom bombardment mass spectrometry; GC/EIMS, gas chromatography/electron impact mass spectrometry; GSL(s) glycosphingolipids; HPTLC, high performance thin-layer chromatography; Neu5Ac,N-acetylneuraminic acid [26], PFU, plaque forming unit. The designation of the following glycosphingolipids follows the IUPAC-IUB recommendations, and the ganglioside nomenclature system of Svennerholm was used. LacCer or lactosylceramide, Gal1-4Glc1-1Cer gangliotetraosylceramide or GgOse4Cer, Gal1-3GalNAc1-4Gal1-4Glc1-1Cer; lacto-N-tetraosylceramide or nLcOse4Cer, Gal1-4GlcNAc1-3Gal1-4-Glc1-1Cer; lacto-N-norhexaosylceramide or nLcOse6Cer, Gal1-4GlcNAc1-3Gal1-4GlcNAc1-3Gal 1-4-Glc1-1Cer; GM3, II3Neu5Ac-LacCer; GM1, II3Neu5Ac-GgOse4Cer; GD1a, IV3Neu5Ac, II3Neu5Ac-GgOse4Cer; GD1b, II3(Neu5Ac)2-GgOse4Cer; GT1b, IV3Neu5Ac, II3(Neu5Ac)2-GgOse4Cer; GQ1b, IV3(Neu5Ac)2, II3(Neu5Ac)2-GgOse4Cer; sialyllacto-N-tetraosylceramide, IV3Neu5Ac/IV6Neu5Ac-nLcOse4Cer; sialyllacto-N-norhexaosylceramide or i-active ganglioside, VI3Neu5Ac-nLcOse6Cer. 相似文献
7.
Harald Rösner 《Development genes and evolution》1980,188(3):205-213
Summary The developmental profiles of 15 different gangliosides of the optic lobes and cerebrum of the chicken were followed from the 6 th day of incubation to hatching and correlated to morphological development. Five of these gangliosides appearing in both structures between the sixth and tenth day, have not been reported previously in higher vertebrates. Three chromatographed on TLC-plates similarly to GT3, GT2, and GT1c gangliosides, which have been demonstrated in fish brain. One fraction moved just below GQ1b and is suggested, to contain GQ1c. These novel gangliosides, which are possibly related to a recently proposed separate and probably phylogenetically older biosynthetic pathway, contained up to 20% of total ganglioside sialic acid. The fifth novel fraction, containing up to 16% of total ganglioside-sialic acid, moved below the penta-sialoganglioside GP1 and is suggested to contain hexa-sialogangliosides.There were two main changes in ganglioside synthesis, which were identical in both structures.The first occurred from the sixth to the eleventh day, parallel to decreased proliferation, maximal cell migration and neuroblast differentiation, GD3 and GD2 decreased rapidly in favour of GQ1b, GP1, and to the novel fractions, described above.The second occurred from the eleventh to the eighteenth day, parallel to increased growth and arborization of dendrites and axons as well as functional establishment of synaptic contacts, there was a sharp rise in the amount of GD1b, GT1b, and GD1a. Concomitantly the novel gangliosides decreased. At hatching GD1a was the predominant ganglioside. GM3, GM2, and GM1 were always minor fractions, each accounting for less than 4% of total ganglioside-sialic acid. GM4 was never detected, indicating neglegible myelinisation until hatching.Abbreviations NeuAc
N-Acetylneuraminic acid
- GM4
I3NeuAc-GalCer
- GM3
II3NeuAc-LacCer
- GM2
II3NeuAc-GgOse3Cer
- GM1
II3NeuAc-GgOse4Cer
- GD3
II3(NeuAc)2LacCer
- GD1a
IV3NeuAc, II3NeuAc GgOse4Cer
- GT3
II3 (NeuAc)3LacCer
- GD2
II3(NeuAc)2GgOse3Cer
- GD1b
II3(NeuAc)2GgOse3Cer
- GD1b
II3(NeuAc)2GgOse4Cer
- GT2
II3(NeuAc)3GgOse3Cer
- GT1b
IV3NeuAc, II3(NeuAc)2GgOse4Cer
- GT1c
II3(NeuAc)3GgOse4Cer
- GQ1b
IV3(NeuAc)2 II3(NeuAc)2GgOse4Cer
- GQ1c
IV3NeuAc, II3(NeuAc)3GgOse4Cer
- GP1
IV3(NeuAc)2, II3(NeuAc)3GgOse4Cer
- GH(?)
IV3(NeuAc)3, II3(NeuAc)3GgOse4Cer 相似文献
8.
Harald Rösner 《Development genes and evolution》1977,183(4):325-335
Summary The developmental accretion of up to nine individual gangliosides in foetal brains, peri- and postnatal cortices, postnatal cerebelli and olfactory lobes and in the liver and the spleen were investigated in mice and compared with that of glycoprotein-bound sialic acid and the activity of the acetylcholinesterase.In foetal brain and in postnatal liver and spleen more sialic acid was found bound to glycoproteins than to gangliosides. In postnatal brain structures, however, ganglioside-NeuAc predominated and increased between the 7th and 21st d about 2-fold in the olfactory lobes and cerebellum and more than 3-fold in the cortex.During foetal development the relative quantities (mol %) as well as the absolute concentrations (compared with the fresh weight) of GM1, GM2 and GM3 in the brain decreased, whereas those of GD1a, GD1b and GQ increased.This pattern change continued perinatally in the cortex up to the end of the first week. Thereafter the pattern changed little, but the concentration of all gangliosides present increased much more rapidly, especially between the 10th and 13th d.The postnatal cerebellum and olfactory lobes contained higher concentrations of GM1 and GM3 than the cortex, both gangliosides decreasing in favour of their di-, tri- and tetrasialo-homologues during the third postnatal week.In all brains structures the accretion of GD1a and GT1 was proportional to the increase in the activity of the acetylcholinesterase.Unlike the brain structures, the ganglioside pattern in the liver and spleen, characterised by a predominance of monosialogangliosides and of GD3, did not change noticeably during the first three weeks after birth.The coincidence of the changes in ganglioside accretion observed in the different brain structures with successive periods of morphological differentiation further support the suggestion that gangliosides may play an important role in control of the growth and differentiation of developing nerve cells.Abbreviations GM3
II3NeuAc-GgOse2Cer
- GM2
II3NeuAc-GgOse3Cer
- GM1
II3NeuAcGgOse4Cer
- GD1a
IV3NeuAc-, II3 NeuAc-GgOse4Cer
- GD3
II3 NeuAc2-GgOse2Cer
- GD2
II3 NeuAc2-GgOse3 Cer
- GD1b
II3 NeuAc2-GgOse4 Cer
- GT1
IV3 NeuAc-, II3 NeuAc2-GgOse4 Cer
- GQ
IV3 NeuAc-, II3 NeuAc3-GgOse4 Cer
- NeuAc
N-acetylneuraminic acid (sialic acid)
- AChE
Acetylcholinesterase 相似文献
9.
EnterotoxigenicEscherichia coli (ETEC) strains expressing F5 (K99) fimbriae cause diarrhoea in the young animal through adhesion to specific sialoglycolipids of the small intestine surface. We studied here an infant mouse diarrhoea model, as CBA infant mice are susceptible to F5-positive ETEC infection, whereas DBA/2 ones are resistant. In an attempt to determine an enzymatic basis for susceptibility and resistance, we investigated the intestine ganglioside pattern in relation to the activity of glycosyltransferases responsible for the globo- and ganglio-series. We observed that the intestine of susceptible CBA infant mice displayed a characteristic sialoglycolipid pattern containing mainly the F5 receptors. The two murine strains differed in the relative activities of galactosyltransferases (GbOse3Cer and GM1 synthases),N-acetylgalactosylaminyltransferases (GA2 and GM2 synthases) and sialyltransferases (GM3 and GD3 synthases). An elevated GM3-synthase activity was observed in the intestine of susceptible CBA infant mice, at the age of high susceptibility. Hence, we conclude that the marked specificity of mouse type correlated with susceptibility and resistance to F5-positive ETEC infection which could be controlled through the regulation of glycosyltransferase activities.Abbreviations NeuAc
N-acetylneuraminic acid
- NeuGc
N-glycolylneuraminic acid
- Glc
glucose
- GalNAc
N-acetylgalactosamine
- Gal
galactose
- Car
ceramide
- LacCer
lactosylceramide (Galß-4Glcß1-1Cer)
- GA2
asialo-GM2 (GgOse3Cer)
- GA1
asialo-GM1 (GgOse4Cer)
- NeuAc/NeuGc-GMla
II3 NeuAc/NeuGc-GgOse4Cer
- NeuAc/NeuGc-GM1a
IV3 NeuAc/NeuGc-GgOse4Cer
- NeuAc/NeuGc-GM2
II3 NeuAc/neuGc-GgOse3Cer
- NeuAc/NeuGc-GM3, II3 NeuAc/NeuGc-LacCer; NeuAc/NeuGc-GD1a, IV3 NeuAc/NeuGc, II3 NeuAc/NeuGc-GgOse4Cer; NeuAc/NeuGc-GD1b
II3 (NeuAc/NeuGc)2-GgOse4Cer
- NeuAc/NeuGc-GD1c
IV3 (NeuAc/NeuGc)2-GgOse4Cer
- NeuAc/NeuGc-GD2, II3 (NeuAc/NeuGc)2-GgOse3Cer; NeuAc/NeuGc-GD3, II3 (NeuAc/NeuGc)2-Lac Cer; NeuAc/NeuGcGT1a
IV3 (NeuAc/NeuGc)2, II3 NeuAc/NeuGc-GgOse4Cer
- NeuAc/neuGc-GT1b
IV3 NeuAc/NeuGc, II3 (NeuAc/NeuGc)2-GgOse4Cer
- NeuAc/NeuGc-GT1c
II3 (NeuAc/NeuGc)3-GgOse4Cer; NeuAc/NeuGc-GT2, II3 (NeuAc/NeuGc)3-GgOse3Cer
- NeuAc/NeuGc-GT3
II3 (NeuAc/NeuGc)3-Lac Cer
- NeuAc/NeuGc-GQ1b
IV3 (NeuAc/NeuGc)2, II3 (NeuAc/NeuGc)2-GgOse4Cer
- NeuAc/NeuGc-GQ1c
IV3 NeuAc/NeuGc, II3 (NeuAc/NeuGc)3-GgOse4Cer
- NeuAc/NeuGc-GP1c
IV3 (NeuAc/NeuGc)2, II3 (NeuAc/NeuGc)3-GgOse4Cer
- GD, GT and GQ
di-, tri- and tetra-sialoglangliosides. NeuGc-SPG, IV3 NeuGc-nLcOse4Cer. Glycosyltransferases assayed in this work areN-acetylgalactosaminyltransferases
- UDP-GalNAc
lactosylceramide 1-4N-acetylgalactosaminyltransferase or GA2 synthase (EC 2.4.1-) and UDP-GalNAc:(N-acetylneuraminyl)-lactosylceramide 1-4N-acetylgalactosaminyltransferase or GM2 synthase (EC 2.4.1.92)
- sialyltransferases
CMP-N-acetylneuraminate: lactosylceramide 2–3 sialyltransferase (sialyltransferases I and IV) or GM3 synthase (EC 2.4.99.-) and CMP-N-acetylneuraminate:(N-acetylneuraminyl) lactosylceramide 2-8 sialyltransferase (sialyltransferase II) or GD3 synthase (EC 24.99.8)
- galactosyltransferases
UDP-galactose:N-acetylgalactosaminyl-(N-acetylneuraminyl) lactosylceramide 1-3 galactosyltransferase (galactosyltransferase II) or GM1a synthase (EC 2.4.1.62) and UDP-galactose:lactosylceramide 1-4 galactosyltransferase or GbOse3Cer synthase (EC 2.4.1-) 相似文献
10.
Ganglioside alterations in YAC-1 cells cultivated in serum-supplemented and serum-free growth medium
Gangliosides of the GM1b-pathway (GM1b and GalNAc-GM1b) have been found to be highly expressed by the mouse T lymphoma YAC-1 grown in serum-supplemented medium, whereas GM2 and GM1 (GM1a-pathway) occurred only in low amounts [Müthing, J., Peter-Katalini, J., Hanisch, F.-G., Neumann, U. (1991)Glycoconjugate J
8:414–23]. Considerable differences in the ganglioside composition of YAC-1 cells grown in serum-supplemented and in well defined serum-free medium were observed. After transfer of the cells from serum-supplemented medium (RPMI 1640 with 10% fetal calf serum) to serum-free medium (RPMI 1640 with well defined supplements), GM1b and GalNAc-GM1b decreased and only low amounts of these gangliosides could be detected in serum-free growing cells. The expression of GM1a was also diminished but not as strongly as that of GM1b and GalNAc-GM1b. These growth medium mediated ganglioside alterations were reversible, and the original ganglioside expression was achieved by readaptation of serum-free growing cells to the initial serum-supplemented medium. On the other hand, a new ganglioside, supposed to represent GalNAc-GD1a and not expressed by serum-supplemented growing cells, was induced during serum-free cultivation, and increased strongly after readaptation. These observations reveal that the ganglioside composition ofin vitro cultivated cells can be modified by the extracellular environment due to different supplementation of the basal growth medium.
Abbreviations: BSA, bovine serum albumin GSL(s), glycosphingolipid(s); HPTLC, high-performance thin-layer chromatography; LDL, low density lipoprotein; NeuAc,N-acetylneuraminic acid; NeuGc,N-glycoloylneuraminic acid. The designation of the following glycosphingolipids follows IUPAC-IUB recommendations. GgOse3Cer or gangliotriaosylceramide, GalNAc1-4Gal1-4GlcCer; GgOse4Cer or gangliotetraosylceramide, Gal1-3GalNAc1-4Gla1-4GlcCer; GgOse5Cer or gangliopentaosylceramide, GalNAc1-4Gal1-3GalNAc1-4Gal1-4GlcCer; GgOse6Cer or gangliohexaosylceramide, Gal1-3GalNAc1-4Gal1-3GalNAc1-4Gal1-4GlcCer or GgOse6Cer; II3NeuAc-GgOse3Cer or GM2; II3NeuAc-GgOse4Cer or GM1 or GM1a; IV3NeuAc-GgOse4Cer or GM1b; IV3NeuAc-GgOse5Cer or GalNAc-GM1b; IV3NeuAc-GgOse6Cer or Gal-GalNAc-GM1b; IV3NeuAc, II3NeuAc-GgOse4Cer or GD1a; II3(NeuAc)2-GgOse4Cer or GD1b; IV3NeuAc, III6NeuAc-GgOse4Cer or GD1a; IV3NeuAc, II3NeuAc-GgOse5Cer or GalNAc-GD1a.
Enzymes: Vibrio cholerae andArthrobacter ureafaciens neuraminidase (EC 3.2.1.18). 相似文献
11.
A rapid procedure is described for the separation of CMP-sialic acid:lactosylceramide sialyltransferase reaction components using Sep Pak C18 cartridges. The quantitative separation of the more polar nucleotide sugar, CMP-sialic acid, and its free acid from the less polar GM3-ganglioside is simple and rapid relative to previously described methods. Recovery of GM3 is optimized by the addition of phosphatidylcholine to the reaction mixture prior to the chromatographic step. Using rat liver Golgi membranes as a source of CMP-sialic acid: lactosylceramide sialyltransferase activity (GM3 synthase; ST-1), the transfer of [14C] sialic acid from CMP-[14C] sialic acid to lactosylceramide can be quantified by this assay. The procedure is reliable and may be applicable to the isolation of ganglioside products in otherin vitro glycosyltransferase assays.Abbreviations GM3
GM3-ganglioside
- II3NeuAc-LacCer
NeuAc2-3Gal1-4Glc1-1Cer
- GD1a
GD1a-ganglioside, IV3NeuAc, II3NeuAc-GgOse4Cer, NeuAc2-3Gal1-3GalNac1-4(NeuAc2-3)Gal1-4Glc1-1Cer
- GD3
GD3-ganglioside, II3(NeuAc)2LacCer, NeuAc2-8NeuAc2-3Gal1-4Glc1-1Cer
- GgOse4Cer
asialo-GM1 Gal1-3GalNAc1-4Gal1-4Glc1-1Cer
- FucGMI
fucosyl-GMI-ganglioside, Fuc1-2Gal1-3GalNAc1-4Gal1-4 Glc1-1Cer
- ST-1
GM3 synthase, CMP-sialic acid:lactosylceramide sialyltransferase
- LacCer
lactosylceramide, Gal1-4Glc1-1Cer
- CMP-NeuAc
cytidine 5-monophospho-N-acetylneuraminic acid
- PC
phosphatidylcholine
- PMSF
phenylmethylsulfonyl fluoride 相似文献
12.
Sialidases cleave off sialic acid residues from the oligosaccharide chain of gangliosides in their catabolic pathway while sialyltransferases transfer sialic acid to the growing oligosaccharide moiety in ganglioside biosynthesis. Ganglioside GM3 is a common substrate for both types of enzymes, for sialidase acting on ganglioside GM3 as well as for ganglioside GD3 synthase. Therefore, it is possible that both enzymes recognize similar structural features of the sialic acid moiety of their common substrate, ganglioside GM3. Based on this idea we used a variety of GM3 derivatives as glycolipid substrates for a bacterial sialidase (Clostridium perfringens) and for GD3 synthase (of rat liver Golgi vesicles). This study revealed that those GM3 derivatives that were poorly degraded by sialidase also were hardly recognized by sialyltransferase (GD3 synthase). This may indicate similarities in the substrate binding sites of these enzymes. 相似文献
13.
Guiliano Gazzotti Sandro Sonnino Riccardo Ghidoni Paolo Orlando Guido Tettamanti 《Glycoconjugate journal》1984,1(2):111-121
Gangliosides GM2, GM1 and GD1b were radiolabelled at C-6 of the terminal galactose orN-acetylgalactosamine by the galactose oxidase/[3H]NaBH4 method; gangliosides GM2, GM1, Fuc-GM1 and GD1a were radiolabelled at C-3 of the long chain base by the 2,3-dichloro-5,6-dicyanobenzoquinone/[3H]NaBH4 method.By application of an original HPLC procedure, eight different molecular species were prepared from each labelled ganglioside. Each of these species was characterized by the presence of one of the following long chain bases:erythro C18 sphingosine,threo C18 sphingosine,erythro C18 sphinganine,threo C18 sphinganine,erythro C20 sphingosine,threo C20 sphingosine,erythro C20 sphinganine andthreo C20 sphinganine.From GD1b only the species containing theerythro forms of long chain bases were obtained.The individual molecular species were more than 99% homogeneous and had a radiopurity better than 99%. The molecular species of the same ganglioside, radiolabelled at C-3 of the long chain base, had identical specific radioactivity, namely 1.17, 1.25, 0.85 and 1.28 Ci/mmol for GM2, GM1, Fuc-GM1 and GD1a respectively. The molecular species of the same ganglioside, radiolabelled at C-6 of terminal galactose orN-acetylgalactosamine, had similar specific radioactivity, namely 1.34–1.40, 1.44–1.51, 1.37–1.44 Ci/mmol for GM2, GM1 and GD1b respectively. 相似文献
14.
Vanna Chigorno Marina Pitto Gloria Cardace Domenico Acquotti Günther Kirschner Sandro Sonnino Riccardo Ghidoni Guido Tettamanti 《Glycoconjugate journal》1985,2(3-4):279-291
The preparation of a GM1-ganglioside (GM1) [14C]-labelled in the sialic acid residue is reported. This can be obtained by re-N-acetylation in the presence of [1-14C]-acetic anhydride, of a GM1 derivative de-N-acetylated specifically on the sialic acid residue by alkaline hydrolysis of GM1 with tetramethylammonium hydroxide. The radiolabelled GM1 is utilized to investigate the binding properties and the mode of interaction of GM1 with cultured fibroblasts. Three different forms of association (one serum-removable, one trypsin-removable and one trypsin-stable) have been recognized to occur in a way that depended on cell culture conditions (presence or absence of fetal calf serum), ganglioside concentration (from, 5×10–9 M to 10–4 M) and incubation time (up to 24 h). Some metabolic modifications of GM1 during the period of high cell viability were also investigated.Abbreviations GM1
GM1-ganglioside, II3NeuAc-GgOse4Cer
- FCS
fetal calf serum
- EMEM
Eaglés Minimum Essential Medium with Earlés salts
- PBS
Dulbecco phosphate buffered saline without calcium and magnesium 相似文献
15.
Gangliosides with a short carbohydrate chain: II3(NeuAc)2-LacCer, II3(NeuAc)2-GgOse3Cer, II3(NeuAc)3-LacCer, II3NeuAc-LacCer, and II3NeuAC-GgOse3 Cer, were found to be predominant in the brain of 8 species of cartilaginous fish, elasmobranches. N-acetylneuraminic acid was the only sialic acid found in these gangliosides, the N-glycolyl derivative being practically absent. 4-Sphingenine was shown to be the predominant sphingoid in elasmobranch brain gangliosides. The sequential enzymatic hydrolysis of II3(NeuAc)2 -LacCer from shark and ray brain by acylneur-aminyl hydrolase (EC 3.2.1.18) and β-D-galactoside-galactohydrolase (EC 3.2.1.23), as well as permethylation studies, provide further evidence for the following structure of this major elasmobranch brain ganglioside: 相似文献
16.
Heike Brandt Johannes Müthing Jasna Peter-Katalinić Jürgen Lehmann 《Cytotechnology》1994,16(2):89-100
Gangliosides of the mouse-rat hybridoma cell line 187.1, which secretes an antibody against -light chain of mouse IgG, were isolated and structurally characterized by biochemical and immunological methods (overlay technique), and fast atom bombardment-mass spectrometry. Exclusively G
M3, substituted with C241 and C160 fatty acid and C181 sphingosine, was found in this B cell derived cell line. A G
M3 (NeuGc) to G
M3(NeuAc) ratio (80 to 20), was characteristic for 187.1 cells, and absolute G
M3 amounts of about 0.3 mg 10–9 viable cells were determined. Exogenous application of G
M3, which has been isolated from large cell preparations, to 187.1 cells showed growth inhibition in a concentration dependent manner. Using the MTT-assay and the [3H]thymidine incorporation assay, the cells exhibited a strong reduction in metabolic and proliferative activity, respectively, after exposure of cells to G
M3. G
M3 was applied in concentrations between 3M and 30M, giving evidence for strong inhibitory effects at 30M G
M3 and less but significant suppression after application of G
M3 concentrations lower than 20M. No cellular response was observed at the lowest concentration (3M) used in this study. Hybridoma cells as well as other cell types like fibroblasts, muscle cells and endothelial cells, are in general characterized by high expression of the G
M3 ganglioside, which is known to act as a modulator of cellular growth in monolayer cultures of adherent cells. Since gangliosides are released to the culture medium by cell lysis, i.e. cell death, and/or by active membrane shedding, the results obtained in this study suggest a growth regulatory role of G
M3 in high density hybridoma cell cultures.Abbreviations DMB
1,2-diamino-4,5-methylenedioxybenzene
- FAB-MS
fast atom bombardment-mass spectrometry
- GSL(s)
glycosphingolipid(s)
- HPLC
high performance liquid chromatography
- HPTLC
high performance thin layer chromatography
- MTT
3,(4,5 dimethylthiazol-2-yl)2,5 diphenyl tetrazolium bromide
- NeuAc
N-acetylneuraminic acid
- NeuGc
N-glycolylneuraminic acid
- PBS
phosphate buffered saline
The designation of the following glycosphingolipids follows the IUPAC-IUB recommendations (1977) and the nomenclature of Svennerholm (1963). Lactosylceramide or LacCer, Galß1–4Glcß1-1Cer; gangliotriaosylceramide or GgOse3Cer; GalNAcß1–4Galß1–4Glcß1-1 Cer; gangliotetraosylceramide or GgOse4Cer, Galß1–3GalNAcß1–4Galß1–4Glcß1-1Cer; G
M3(NeuAc), II3NeuAc-LacCer; G
M3(NeuGc), II3NeuGc-LacCer; G
M2(NeuGc), II3NeuGc-GgOse3Cer; G
M1 or G
M1a, II3NeuAc-GgOse4Cer; G
M1b, IV3NeuAc-GgOse4Cer. 相似文献
17.
Kenneth C. Leskawa Robert E. Erwin Paul E. Buse Edward L. Hogan 《Molecular and cellular biochemistry》1988,83(1):47-54
Summary Glycosphingolipid biosynthesis was examined using [3H]-galactose as a precursor as rat L6 myoblasts fused to form multinucleated myotubes. Incorporation of label into neutral glycolipids decreased steadily as the population of myotubes increased, so that final biosynthesis was one-half that observed with myoblasts (p < 0.02). Conversely, ganglioside biosynthesis doubled during myoblast confluency (p < 0.02) and then decreased as myotubes formed. Qualitatively, L6 cells synthesized large amounts of ganglioside GM3 during all myogenic phases. The major neutral glycosphingolipid products were lactosylceramide and paragloboside (nLcOse4Cer). Few changes in TLC autoradiographic patterns were noted during differentiation, with the exception of a slight decrease in ganglioside GM1. The results indicate that the biosynthesis of glycosphingolipids is tightly regulated during myogenesis in vitro and suggest a role for membrane gangliosides in muscle cell differentiation.Abbreviations GM1
II3NeuAc-GgOse4Cer
- GM3
II3NeuAc-GgOse2Cer
- MG4
IV3NeuAc-nLcOse4Cer
- MG6
VI3NeuAc V4Gal-IV3GlcNAc-nLcOse4Cer
- TLC
Thin-Layer Chromatography
- DMEM
Dulbecco's Modified Eagles' Medium 相似文献
18.
Dorothee Heuermann Peter Roggentin Reinhard G. Kleineidam Roland Schauer 《Glycoconjugate journal》1991,8(2):95-101
The sialidase secreted byClostridium chauvoei NC08596 was purified to apparent homogeneity by ion-exchange chromatography, gel filtration, hydrophobic interaction-chromatography, FPLC ion-exchange chromatography, and FPLC gel filtration. The enzyme was enriched about 10 200-fold, reaching a final specific activity of 24.4 U mg–1. It has a relatively high molecular mass of 300 kDa and consists of two subunits each of 150 kDa. The cations Mn2+, Mg2+, and Ca2+ and bovine serum albumin have a positive effect on the sialidase activity, while Hg2+, Cu2+, and Zn2+, chelating agents and salt decrease enzyme activity. The substrate specificity, kinetic data, and pH optimum of the enzyme are similar to those of other bacterial sialidases.Abbreviations FPLC
fast protein liquid chromatography
- NCTC
National Collection of Type Cultures
- ATCC
American Type Culture Collection
- MU-Neu5Ac
4-methylumbelliferyl--d-N-acetylneuraminic acid
- buffer A
0.02m piperazine, 0.01m CaCl2, pH 5.5
- buffer B
0.02m piperazine, 0.01m CaCl2, 1.0m NaCl, pH 5.5
- buffer C
0.1m sodium acetate, 0.01m CaCl2, pH 5.5
- SDS
sodium dodecyl sulfate
- PAGE
polyacrylamide gel electrophoresis
- Neu5Ac
N-acetylneuraminic acid
- BSM
bovine submandibular gland mucin
- GD1a
IV3Neu5Ac, II3Neu5Ac-GgOse4Cer
- GM1
II3Neu5Ac-GgOse4Cer
- MU-Neu4,5Ac2
4-methylumbelliferyl--d-N-acetyl-4-O-acetylneuraminic acid
- TLC
thin-layer chromatography
- HPTLC
high performance thin-layer chromatography
- EDTA
ethylenediamine tetraacetic acid
- EGTA
ethylene glycol bis(2-aminoethyl-ethen)-N,N,N,N-tetraacetic acid
- BSA
bovine serum albumin
- Neu5Ac2en
2-deoxy-2,3-didehydro-N-acetylneuraminic acid
- IEF
isoelectric focusing
- IEP
isoelectric point 相似文献
19.
Yasuhiro Hashimoto Mikiko Abe Akemi Suzuki Kentaro Iwasaki Tamio Yamakawa 《Glycoconjugate journal》1985,2(3-4):255-265
Genetic polymorphism in the expression of the GM1(NeuGc) ganglioside has been shown in the liver of inbred strains of mice. Through analysis of the gangliosides of H-2 congenic and recombinant strains, this polymorphism was demonstrated to be controlled by a locus mapped left outside of the H-2 complex on chromosome 17, and the locus was assumed to control the level of the activity of GM1(NeuGc) synthetase, UDP-galactose:GM2(NeuGc) galactosyltransferase (E.C.2.4.1.62) [Hashimotoet al., J Biochem (1983) 94:2049-54].In the present study we analyzed the genetic linkage between the activity of the galactosyltransferase and the H-2 haplotype. For this purpose, we selected two inbred strains of mice, WHT/Ht and BALB/c, because they have different levels of the transferase activity and show different H-2 haplotypes; the specific activity of the transferase obtained with BALB/c was one-eighth of that with WHT/Ht, and BALB/c expressed the la.7 antigen as one of the products encoded in their H-2d complex, whereas WHT/Ht did not. To analyze the linkage between these two phenotypes, WHT/Ht were mated with BALB/c to obtain the F1 mice, and the female F1 mice were then backcrossed to WHT/Ht. It was found that one half of the backcross generation expressed the la.7 antigen derived from BALB/c and had a significantly lower specific activity of the transferase than that of WHT/Ht, while the other half did not express the la.7 antigen but had the same specific activity of the transferase as that obtained with WHT/Ht.These results suggest that the locus controlling the level of the transferase activity in mouse liver is linked to the H-2 complex on chromosome 17.Abbreviations NeuGc
N-glycolylneuraminic acid
The ganglioside nomenclature is based on the system of Svennerholm, J Neurochem (1963) 10:613-23. The sialic acid species present is shown in parentheses after the ganglioside abbreviation. 相似文献
20.
Kentaro Horikawa Masahiko Yamasaki Masao Iwamori Hideki Nakakuma Kiyoshi Takatsuki Yoshitaka Nagai 《Glycoconjugate journal》1991,8(4):354-360
We analysed the glycolipids of mouse thymocytes before and after Concanavalin A (Con A) or recombinant interleukin-2 (rIL-2) stimulation by TLC-immunostaining with carbohydrate-specific antiglycolipid antibodies. The thymocytes were cultured in serum-free medium in the presence of 500 ng ml–1 Con A, 10 U ml–1 rIL-2 or Con A plus rIL-2 for 6, 12, 24, 48, and 72 h, and were found to start proliferating 24 h after cultivation in the presence of Con A or Con A plus rIL-2, the maximum levels being reached at 72 h and 48 h, respectively, in a thymidine uptake experiment. The concentrations of II3Neu-Gg4Cer, Gg4Cer and IV3GalNAc-Gb4Cer after 48 h Con A stimulation were found to be at almost the original levels. Conversely, II3Neu-Gg3Cer, which was not detected in the thymocytes at the start, began to appear after 48 h stimulation with Con A and Con A plus rIL-2, and IV3Neu-Gg5Cer in the cells 48 h after stimulation with Con A and Con A plus rIL-2 has increased to 41 and 44 times higher than in the original cells, respectively, as judged on TLC-immunostaining with monoclonal antibody YHD-06, which detects the GalNAc1-4(NeuAc or NeuGc2-3)Gal-structure. These results indicate that the increased synthesis of both gangliosides, in other words, the activation ofN-acetylgalactosaminyltransferase, is associated with the mitogen-induced proliferation.N-Acetylneuraminic acid was the sole sialic acid in II3Neu-Gg3Cer which newly appeared in the cells on stimulation, whereas the sialic acid of IV3Neu-Gg5Cer was a mixture ofN-acetyl- andN-glycoloylneuraminic acids. This result may suggest that the substrates for the two differentN-acetylgalactosaminyltransferases must be different. This GalNAc1-4(NeuAc or NeuGc2-3)Gal-structure was also detected on the surface of the Con A or Con A plus rIL-2 stimulated mouse thymocytes on flow cytometric analysis of cells indirectly stained with monoclonal antibody YHD-06.
Abbreviations: carbohydrate and glycolipid nomenclature and abbreviations follow the IUPAC-IUB recommendations or the nomenclature system of Svennerholm L. (1963)J Neurochem
10:613–23. 相似文献