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1.
Daisei Miyamoto Takao Ueno Sachiko Takashima Kazuhide Ohta Toshio Miyawaki Takashi Suzuki Yasuo Suzuki 《Glycoconjugate journal》1997,14(3):379-388
A new monoclonal antibody (TU-1) directed against the Galα1-4Galβ1-4Glc residue of the Gb3Cer/CD77 antigen was prepared by
the hybridoma technique following immunization of mice with an emulsion composed of monophosphoryl lipid A, trehalose dimycolate,
and Gb3Cer isolated from porcine erythrocytes. TU-1 showed reactivity towards Gb3Cer and lyso-Gb3Cer (Galα1-4Galβ1-4Glcβ1-1′Sph),
although the reactivity towards lyso-Gb3Cer was about 10-fold lower than that to Gb3Cer. But it did not react with other structurally-related
glycolipids, such as LacCer (Galβ1-4Glcβ1-1′Cer), Gg3Cer, Gg4Cer, Gb4Cer (GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1′Cer), galactosylparagloboside
(Galα1-3Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1′Cer), sulfatide (HSO3-3Galβ1-1′Cer), other gangliosides (GM3, GM2, GM1a, GD1a and
GT1b), or P1 antigen (Galα1-4Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1′Cer) among neutral glycolipids prepared from P1 phenotype red
blood cells. Furthermore, TU-1 reacted with viable lymphoma cells, such as human Burkitt lymphoma cell line, Daudi, and Epstein-Barr
virus (EBV)-transformed B cells by the immunofluorescence method, and also with germinal centre B cells in human tonsil and
vessel endothelial cells in human thymus histochemically. These results indicate that TU-1 is a monoclonal antibody directed
against Gb3Cer/CD77 antigen and can be utilized as a diagnostic reagent for Burkitt's lymphoma and also for detection of the
blood group Pk antigen in glycolipid extracts of erythrocytes. Abbreviations: ATL, adult T-cell leukaemia; BSA, bovine serum
albumin; Cer, ceramide; DPPC, L-α-dipalmitoylphosphatidylcholine; EBV, Epstein-Barr virus; FCS, fetal calf serum; GalCer,
Galβ1-1′Cer; GlcCer, Glcβ1-1′Cer; LacCer, Galβ1-4Glcβ1-1′Cer; Gb3Cer, Galα1-4Galβ1-4Glcβ1-1′Cer; Iyso-Gb3Cer, Galα1-4Galβ1-4Glc1-1′Sph;
Gb4Cer, GalNAcβ1-3Galα1-4Galβ1-4Glc1-1′Cer; galactosylparagloboside, Galα1-3Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1′Cer; Gg3Cer, GalNAcβ1-4Galβ1-4Glcβ1-1′Cer;
Gg4Cer, Galβ1-3GalNAcβ1-4Galβ1-4Glcβ1-1′Cer; GM3, Neu5Acα2-3Galβ1-4Glcβ1-1′Cer; GM2, GalNAcβ1-4(Neu5Acα2-3) Galβ1-4Glcβ1-1′Cer;
GM1a, Galβ1-3GalNAcβ1-4(Neu5Acα2-3)Galβ1-4Glcβ1-1′Cer; GD1a, Neu5Acα2-3Galβ1-3GalNAcβ1-4(Neu5Acα2-3)Galβ1-4Glcβ1-1′Cer; GD1b,
Galβ1-3GalNAcβ1-4(Neu5Acα2-8Neu5Acα2-3)Galβ1-4Glcβ1-1′Cer; GT1b, Neu5Acα2-3Galβ1-3GalNAcβ1-4(Neu5Acα2-8Neu5Acα2-3) Galβ1-4Glcβ1-1′Cer;
HRP, horseradish peroxidase; LDH, lactate dehydrogenase; MAb, monoclonal antibody; MPL, monophosphoryl lipid A; P1 antigen,
Galα1-4Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1′Cer; PVP, polyvinylpyrolidone; Sph, sphingosine; sulfatide, HSO3-Galβ1-1′Cer; TDM,
trehalose dimycolate; TLC, thin-layer chromatography
This revised version was published online in November 2006 with corrections to the Cover Date. 相似文献
2.
Seven analogues of p-nitrophenyl T-antigen [Galβ(1→3)GalNAcα(1→O)PNP] have been synthesized as potential substrates for elucidation of the substrate specificity of endo-α-N-acetylgalactosaminidase. These compounds, which are commercially unavailable, include: GlcNAcβ(1→3){GlcNAcβ(1→6)}GalNAcα(1→O)PNP [core 4 type], GalNAcα(1→3)GalNAcα(1→O)PNP [core 5 type], GlcNAcβ(1→6)GalNAcα(1→O)PNP [core 6 type], GalNAcα(1→6)GalNAcα(1→O)PNP [core 7 type], Galα(1→3)GalNAcα(1→O)PNP [core 8 type], Glcβ(1→3)GalNAcα(1→O)PNP and GalNAcβ(1→3)GalNAcα(1→O)PNP. The assembly of these synthetic probes was accomplished efficiently, based on di-tert-butylsilylene(DTBS)-directed α-galactosylation as a key reaction. 相似文献
3.
Johannes Muthing 《Glycoconjugate journal》1997,14(2):241-248
The gangliosides GM1b, GalNAc-GM1b and GD1α are typical compounds of concanavalin A stimulated splenic T lymphoblasts of CBA/J
inbred mice. Their structural characterization has been described in previous studies. The intention of this work was the
comparative TLC immunostaining analysis of the glycosphingolipid composition of lectin stimulated splenic T lymphoblasts obtained
from six genetically different inbred mouse strains. The strains examined were AKR, BALB/c, C57BL/6, CBA/J, DBA/2 and WHT/Ht,
which are commonly used for biochemical and immunological studies. The neutral glycosphingolipid GgOse4Cer, the precursor
for GM1b-type gangliosides, was expressed by all six strains investigated. AKR, C57BL/6 and DBA/2 showed high and BALB/c,
CBA/J and WHT/Ht diminished expression in T lymphoblasts, based on single cell calculation. The gangliosides GM1b and GalNAc-GM1b,
elongation products of GgOse4Cer, displayed strain-specific differences in their intensities, which were found to correlate
with the intensities of GgOse4Cer expression of the same strains. Concerning sialic acid substitution of gangliosides, GM1b
and GalNAc-GM1b predominantly carry N-acetylneuraminic acid, whereas choleragenoid receptors GM1a and Gal-GalNAc-GM1b, which
are also expressed by all six strains, are characterized by dominance of N-glycolylneuraminic acid. Two highly polar gangliosides, designated with X and Y, which have not been previously recognized
in murine lymphoid tissue, were detected by positive anti-GalNAc-GM1b antibody and choleragenoid binding, respectively. Both
gangliosides were restricted to AKR, DBA/2 and C57BL/6 mice. The other three strains BALB/c, CBA/J and WHT/Ht are lacking
these structures. In summary, the GM1b-type pathway is quite active in all six strains analysed in this study. Strain-specific
genetic variations in T lymphoblast gangliosides were observed with the occurrence of gangliosides X and Y. This study and
data from other groups strongly indicate for GM1b-type gangliosides a functional association with T cell activation and leukocyte
mediated reactions. Abbreviations: ConA, concanavalin A; GSL(s), glycosphingolipid(s); HPTLC, high-performance thin-layer
chromatography; NeuAc, N-acetylneuraminic acid; NeuGc, N-glycolylneuraminic acid. The designation of the following glycosphingolipids
follows the IUPAC-IUB recommendations (1977) [48] and the ganglioside nomenclature system of Svennerholm [49] for GM1a-type
gangliosides. Glucosylceramide or GlcCer, Glcβ1-1Cer; lactosylceramide or LacCer, Galβ1-4Glcβ1-1Cer; gangliotriaosylceramide
or GgOse3Cer or Gg3, GalNAcβ1-4Galβ1-4Glcβ1-1Cer; gangliotetraosylceramide or GgOse4Cer or Gg4, Galβ1-3GalNAcβ1-4Galβ1-4Glcβ1-1Cer;
gangliopentaosylceramide or GgOse5Cer, GalNAcβ1-4Galβ1-3GalNAcβ1-4Galβ1-4Glcβ1-1Cer; gangliohexaosylceramide or GgOse6Cer,
Galβ1-3GalNAcβ1-4Galβ1-3GalNAcβ1-4Galβ1-4Glcβ1-1Cer. GM3, II3NeuAc-LacCer; GM1 or GM1a, II3NeuAc-GgOse4Cer; GM1b, IV3NeuAc-GgOse4Cer;
GalNAc-GM1b, IV3NeuAc-GgOse5Cer; GD1a, IV3NeuAc, II3NeuAc-GgOse4Cer; GD1b, II3(NeuAc)2-GgOse4Cer; GD1c, IV3(NeuAc)2-GgOse4Cer;
GD1α, IV3NeuAc, III6NeuAc-GgOse4Cer. Only NeuAc-substituted gangliosides are presented in this list of abbreviations
This revised version was published online in November 2006 with corrections to the Cover Date. 相似文献
4.
Cappello S Liu NX Musselli C Brezicka FT Livingston PO Ragupathi G 《Cancer immunology, immunotherapy : CII》1999,48(9):483-492
Fucosyl-GM1 (Fuc-GM1) [Fucα1 → 2Galβ1 → 3GalNAcβ1 → 4(NeuAcα2-3)Galβ1 → 4Glcβ1 → O-Cer] is a small-cell-lung-cancer (SCLC)-associated ganglioside initially defined by the murine monoclonal antibody F12. On
the basis of its known distribution, Fuc-GM1 is a potential target for active immunotherapy in SCLC patients. Fuc-GM1 has
been extracted and purified from bovine thyroid. The immunogenicity of Fuc-GM1 was tested in mice either alone, mixed with
carrier protein keyhole limpet hemocyanin (KLH) or covalently linked with KLH, plus immunological adjuvant QS-21. The Fuc-GM1-KLH
conjugate plus QS-21 adjuvant was found to be optimal. It induced consistent IgM and IgG enzyme-linked immunosorbent assay
(ELISA) titers against Fuc-GM1. These antibodies were strongly reactive with the strongly Fuc-GM1-positive rat hepatoma cell
line H4-II-E, and they were moderately reactive with the moderately positive human SCLC cell line H146 by flow cytometry and
complement-mediated lysis. Both ELISA and fluorescence-activated cell sorting (FACS) reactions were inhibited with Fuc-GM1or
H4-II-E but not with the structurally related ganglioside GM1 or Fuc-GM1-negative colon cancer cell line LS-C. On the basis
of these results, a vaccine comprising Fuc-GM1-KLH plus QS-21 is being prepared for testing in patients with SCLC.
Received: 25 March 1999 / Accepted: 5 August 1999 相似文献
5.
L. A. Simeoni N. E. Byramova N. V. Bovin 《Russian Journal of Bioorganic Chemistry》2000,26(3):183-191
The first synthesis of the Neu5Gc analogue of SiaT
n
disaccharide, which can be detected in breast tumors by immunochemical methods, is reported. The regioselective sialylation
of (3-trifluoroacetamidopropyl)-2-azido-2-deoxy-α-D-galactopyranoside with peracetate of the methyl ester ofN-acetoxyacetyl-neuraminic acid β-ethylthioglycoside in the presence ofN-iodosuccinimide and trifluoromethanesulfonic acid (or its trimethylsilyl ester) resulted in the derivatives of α- and β-sialyl(2→6)galactosaminide
in 39 and 32% yields, respectively. The catalytic hydrogenolysis of the azido group and subsequentN- andO-acetylation of the α-anomer gave the peracetate of trifluoroacetamidopropyl glycoside. Removal of the protective groups led
to glycoside Neu5Gcα2→6GalNAcα-O(CH2)3NH2. Using the Neu5Gc derivative with acetoxyacetyl groups at positions O9 and O4 as a donor increases the α-selectivity of sialylation
to afford the α- and β-anomers in 69 and 8% yields, respectively. 相似文献
6.
Yoon SJ Nakayama K Takahashi N Yagi H Utkina N Wang HY Kato K Sadilek M Hakomori SI 《Glycoconjugate journal》2006,23(9):639-649
GM3 ganglioside interacts specifically with complex-type N-linked glycans having multivalent GlcNAc termini, as shown for
(1) and (2) below. (1) Oligosaccharides (OS) isolated from ConA-non-binding N-linked glycans of ovalbumin, whose structures
were identified as penta-antennary complex-type with bisecting GlcNAc, having five or six GlcNAc termini (OS B1, B2), or bi-antennary
complex-type having two GlcNAc termini (OS I). OS I is a structure not previously described. (2) Multi-antennary complex-type
N-linked OS isolated from fetuin, treated by sialidase followed by β-galactosidase, having three or four GlcNAc termini exposed.
These OS, conjugated to phosphatidylethanolamine (PE), showed clear interaction with 3H-labeled liposomes containing GM3, when various doses of OS-PE conjugate were adhered by drying to multi-well polystyrene
plates. Interaction was clearly observed only with liposomes containing GM3, but not LacCer, Gb4, or GalNAcα1-3Gb4 (Forssman
antigen). GM3 interaction with PE conjugate of OS B1 or B2 was stronger than that with PE conjugate of OS I. GM3 interacted
clearly with PE conjugate of N-linked OS from desialylated and degalactosylated fetuin, but not native fetuin. No binding
was observed to cellobiose-PE conjugate, or to OS-PE conjugate lacking GlcNAc terminus. Thus, GM3, but not other GSL liposomes,
interacts with various N-linked OS having multiple GlcNAc termini, in general. These findings suggest that the concept of
carbohydrate-to-carbohydrate interaction can be extended to interaction of specific types of N-linked glycans with specific
GSLs. Natural occurrence of such interaction to define cell biological phenomena is under investigation.
All solvent ratios are by volume.
An erratum to this article can be found at 相似文献
7.
Peter R. Andreana Przemyslaw Kowal Adam J. Janczuk Peng George Wang 《Glycoconjugate journal》2003,20(2):107-118
Galactose oxidase (EC 1.1.3.9, GAO) was used to convert the C-6′ OH of Galβ(1 → 4)Glcβ–OBn (5) to the corresponding hydrated
aldehyde (7). Chemical modification, through dehydratative coupling and reductive amination, gave rise to a small library
of Galβ(1 → 4)Glcβ–OBn analogues (9a–f, 10, 11). UDP-[6-3H]Gal studies indicated that α1,3-galactosyltransferase recognized the C-6′ modified Galβ(1 → 4)Glcβ–OBn analogues (9a–f,
10, 11). Preparative scale reactions ensued, utilizing a single enzyme UDP-Gal conversion as well as a dual enzymatic system
(GalE and α1,3GalT), taking full advantage of the more economical UDP-Glc, giving rise to compounds 6, 15–22. Galα(1 → 3)Galβ(1
→ 4)Glcβ–OBn trisaccharide (6) was produced on a large scale (2 g) and subjected to the same chemoenzymatic modification as
stated above to produce C-6″ modified derivatives (23–30). An ELISA bioassay was performed utilizing human anti-αGal antibodies
to study the binding affinity of the derivatized epitopes (6, 15–30). Modifications made at the C-6′ position did not alter
the IgG antibody's ability to recognize the unnatural epitopes. Modifications made at the C-6″ position resulted in significant
or complete abrogation of recognition. The results indicate that the C-6′ OH of the αGal trisaccharide epitope is not mandatory
for antibody recognition. Published in 2004.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
8.
GM2 activator protein (GM2AP) is a cofactor for stimulating the enzymatic hydrolysis of the glycolipid GM2 by -hexosaminidase A to produce GM3. We have examined the conformation of GM2AP before and after its interaction with GM2, GM3, and GA2 using circular dichroism and fluorescence spectroscopy techniques. In the presence of GM2, a blue shift of the fluorescence emission maximum and a strong decrease of molar ellipticity values in circular dichroism spectra were observed only at pH 4.5 and at GM2/GM2AP molar ratio higher than 10:1 (up to 50:1). These results suggest that GM2AP assumed a more organized -helical conformation with the tryptophan residues moving from the polar medium toward the hydrophobic environment of the protein. The conformation of GM2AP in the presence of the downstream reaction product, GM3, or a less favorable substrate, GA2, clearly differed from that in the presence of GM2. The relationships between spectroscopic changes and enzymatic activity, herein discussed, strongly suggest that the specific conformation exhibited by GM2AP in the presence of GM2 is functional to serve as an activator for the enzymatic hydrolysis of GM2. 相似文献
9.
Ryszard Russa Maud Bruneteau Alexander S. Shashkov Teresa Urbanik-Sypniewska Hubert Mayer 《Archives of microbiology》1996,165(1):26-33
Lipopolysaccharides from the Rhizobium meliloti wild-type strain 102F51, which is effective in symbiosis with alfalfa, and from the nonnodulating mutant WL113, defective
in root hair adhesion, derived thereof, were isolated and comparatively analyzed. Both preparations were composed of galactose,
glucose, glucuronic acid, galacturonic acid, glucosamine, 3-deoxyheptulosaric acid, and 2-keto-3-deoxyoctonic acid as the
major sugar constitutents. After a modified methylation analysis (consisting of the following consecutive steps: methylation,
carboxyl reduction, remethylation, mild acid hydrolysis, reduction, and trideuterio-methylation), all of the 3-deoxyheptulosaric
and some of the 2-keto-3-deoxyoctonic acid residues were converted into their corresponding 3-deoxyalditol derivatives, which
carried trideuteriomethyl groups at positions C-2, C-4, and C-6. Another part of the permethylated 3-deoxyoctitol was also
found as 2,5,6- and 2,6,8-tri-O-trideuteriomethyl derivatives. NMR data obtained with the separated oligosaccharides and the results of methylation analysis
indicated that the majority of 2-keto-3-deoxyoctonate was present in the fraction of permethylated disaccharide alditols,
namely as 6-O-CD3-aGlc(1→5)3-deoxyoctitol, 6-O-CD3-βGlcNMeAcyl(1→4)3-deoxyoctitol, and as the permethylated trisaccharide alditol, αGalA(1→3)-[6-O-CD3]-β-Glc(1→5)-[4-O-CD3]-3-deoxyoctitol. The presence of trideuteriomethyl groups at C-4 of both 3-deoxyalditols and at C-6 of the glucosaminyl or
glucosyl residues indicated the linkage points of the released acid-labile ketosidic substituents, such as 3-deoxyheptulosarate
and 2-keto-3-deoxyoctonate, in these oligosaccharides. The main differences between the preparations from the wild-type 102F51
and its mutant strain WL 113 were found in the higher content (in strain 102F51) of the following oligosaccharides: α-glucuronosyl(1→4)2-keto-3-deoxyoctonate
and α-galacturonosyl-(1→3)α-glucosyl-(1→5)2-keto-3-deoxyoctonate and in the decreased content of β-glucosaminyl(1→4)2-keto-3-deoxy-octonate.
Received: 21 July 1995 / Accepted: 25 October 1995 相似文献
10.
Perspectives on the significance of altered glycosylation of glycoproteins in cancer 总被引:13,自引:0,他引:13
No abstract Abbreviations: Sia, sialic acid, type unspecified; Tn antigen, GalNAcα 1-O-Ser/Thr; T antigen, Galβ1-3GalNAcα-O-Ser/Thr;
Sialyl LewisX, Siaα2-3Galβ1-4(Fucα1-3)GlcNAc; Sialyl Lewisa, Siaα2-3Galβ1-3(Fucα1-4)GlcNAc; Sialyl-Tn antigen, Siaα2-6GalNAcα1-O-Ser/Thr;
FucT, fucosyltransferase; ST, sialyltransferase.
This revised version was published online in November 2006 with corrections to the Cover Date. 相似文献
11.
Lactococcus lactis subspecies cremoris SBT 0495 produces the phosphopolysaccharide viilian, which consists of the repeating unit β-d-glucosyl-(1→4)-(α-l-rhamnosyl-(1→2))-(α-d-galactose-1-phosphoryl-(→3)-β-galactosyl-(1→4)-β-d-glucose. A lipid extract was prepared from cells in the late exponential phase of growth and was hydrolyzed by hydrochloric
acid under mild conditions to split lipid-linked intermediates in the extract into lipid and sugar moieties. Both moieties
were purified by chromatographic techniques and were characterized to identify intermediates of the viilian biosynthetic pathway.
A polyisoprenoid isolated from the chloroform-soluble fraction of the hydrolyzed lipid extract was identified by mass spectrometry
as undecaprenol. Saccharides isolated from the water-soluble fraction of the hydrolyzed lipid extract by anion-exchange chromatography,
were characterized by glycosidic linkage analysis to discriminate sugar moieties of intermediates of viilian biosynthesis
from compounds liberated from cell wall components. Some oligosaccharide analogues contain a glycerol residue, suggesting
that these are fragments of glycosylglycerides and/or lipoteichoic acid. Three fragments were identified to be glucose, galactosyl-(1→4)-glucose,
and rhamnosyl-(1→2)-galactosyl-(1→4)-glucose, which are in agreement with the structure of the repeating unit of viilian.
These saccharides most likely represent the first three steps of the sequential assembly of the repeating unit of the undecaprenol
assembly.
Received: 2 November 1998 / Accepted: 3 March 1999 相似文献
12.
E.P. Smorodin O.A. Kurtenkov B.L. Sergeyev G.V. Pazynina N.V. Bovin 《Glycoconjugate journal》2003,20(2):83-89
The TF, Tn, and SiaTn glycotopes are frequently expressed in cancer-associated mucins. Antibodies to these glycotopes were
found in human serum. A set of polyacrylamide (PAA)—based glycoconjugates was applied to the direct and competitive enzyme-linked
immunosorbent assays (ELISA) to characterize the specificity of serum IgG antibodies. The anti-TF, -Tn and -SiaTn IgG were
affinity purified from serum of cancer patients and characterized using PAA-conjugates and free saccharides. The anti-TF and
-Tn antibodies were shown to be specific. The anti-TF IgG bound both Galβ1-3GalNAcα- and Galβ1-3GalNAcβ-PAA, the latter was
three-four times more effective inhibitor of antibody binding. The anti-Tn IgG reacted only with GalNAcα-PAA. The anti-SiaTn
IgG cross-reacted with Tn-PAA but SiaTn-PAA was five-six times more effective inhibitor in a competitive assay. The IC50 values for PAA-conjugates with the corresponding antibodies typically ranged from 2 to 5 × 10−8 M. The antibodies display a low specificity to mucin-type glycoconjugates in comparison with PAA-conjugates as was shown
for mucins isolated from human malignant tumor tissues, ovine submaxillary mucin (OSM) and asialo-OSM. The unusual IgG-antibody
specificity to GalNAcβ and GalNAcβ1-3GalNAcβ ligands was found in human serum. Published in 2004.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
13.
N. V. Belyakova O. K. Legina N. L. Ronzhina I. V. Shevelev V. M. Krutiakov 《Biology Bulletin》2010,37(5):464-470
The possibility of interaction of recombinant proteins of human repair DNA polymerase β with proofreading 3′ → 5′-exonucleases
TREX1 and TREX2 was investigated in vitro for the first time. The results of gel filtration analysis show the formation of
a complex between 3′ → 5′-exonucleases mTREX1 and hTREX2 and DNA polymerase β. DNA polymerase activity is shown to increase
four-fold in the presence of 3′ → 5′-exonuclease TREX2. The experiments with the use of immunodot and Western blot assays
on the binding of DNA-polymerase β with 3′ → 5′-exonucleases TREX1 and TREX2 immobilized on a nitrocellulose membrane provided
additional evidence on the direct association of the above proteins in complexes. 相似文献
14.
Akemi Suzuki Shigemi Yoshioka Michiko Sekine Hiromichi Yonekawa Masaru Takenaka Reiji Kannagi 《Glycoconjugate journal》2003,20(3):151-156
The expression of glycan chains is precisely regulated in a time- and space-dependent manner. We summarize here our recent
work on the kidney tubular cell-specific regulation of core 2 β-1,6-GlcNAc transferase. Gsl5 gene was first identified by genetic analysis on the basis of polymorphic expression of kidney glycolipids among inbred strains
of mice and turned out to be a regulatory gene controlling the level of mRNA of kidney-specific core 2 β-1,6-GlcNAc transferase.
This kidney-specific core 2 GlcNAc transferase takes glycolipids having Galβ1-3GalNAc at their termini, Galβ1-3GalNAcα1- and
β1-oligosaccharide derivatives, and glycoproteins having core 1 structure, as substrates. Immunohistochemistry with anti-core
2-Le
x
monoclonal antibody demonstrated that vesicles located just below the microvillous membrane of proximal tubule cells were
clearly stained in a Gsl5-wild type mouse. Western blotting with the monoclonal antibody detected a major glycoprotein with a molecular mass of 500
kDa in the microsomal fraction of the wild type mouse kidney. In situ hybridization with anti-sense cDNA of kidney-specific core 2 GlcNAc transferase confirmed that Gsl5 gene controls the expression of the core 2 β-1,6-GlcNAc transferase mRNA in a proximal tubular cell-specific manner. The
5′ upstream sequences of the kidney-specific core 2 GlcNAc transferase gene in inbred and wild-derived strains of mice were
analyzed, and the phylogenetic analysis of these sequences suggests that functional Gsl5 gene might be produced by the time of subspeciation of M. musculus, about one million years ago. Published in 2004.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
15.
Rare polyagglutinable NOR erythrocytes contain unusual globoside extention products terminating with a Galα1-4GalNAcβ1-3Gal-
unit. This trisaccharide epitope is recognized by recently characterized antibodies naturally occurring in most human sera
(Duk et al., Glycobiology, 15, 109, 2005). These antibodies represent two major types of fine specificity. All these antibodies are most strongly inhibited by Galα1-4GalNAcβ1-3Gal
(NOR-tri), and weakly by Galα1-4Gal. However, the type 1 antibodies are strongly inhibited by Galα1-4Galβ1-3Gal-R and weakly
by Galα1-4GalNAc, while the type 2 antibodies show the opposite reactivities with these two oligosaccharides. Similar antibodies
have now been found in horse, rabbit and pig sera. The antibodies were purified from animal sera by affinity chromatography
on Galα1-4GalNAcβ1-3Gal-human serum albumin(HSA)-Sepharose 4B conjugate. The specificity of the antibodies was determined
by binding to ELISA plates coated with several α-galactosylated oligosaccharide-polyacrylamide (PAA) or -HSA conjugates and
by inhibition with synthetic oligosaccharides. The purified antibodies bound specifically to conjugates containing NOR-tri.
The inhibition of binding showed that the animal sera also contain two types of anti-NOR antibodies: type 2 was found in the
horse serum, and a mixture of both types was present in rabbit and pig serum. These results indicate that anti-NOR, a new
and distinct kind of anti-αGal antibody, are present in animal sera and show similar specificties and diversity as their counterparts
found in human sera. 相似文献
16.
Masłyk M Kochanowicz E Zieliński R Kubiński K Hellman U Szyszka R 《Molecular and cellular biochemistry》2008,312(1-2):61-69
Since Svf1 is phosphoprotein, we investigated whether it was a substrate for protein kinase CK2. According to the amino acid
sequence Svf1 harbours 20 putative CK2 phosphorylation sites. Here, we have reported cloning, overexpression, purification
and characterization of yeast Svf1 as a substrate for three forms of yeast CK2. Svf1 serves as a substrate for both the recombinant
CK2α (K
m 0.35 μM) and CK2α′ (K
m 0.18 μM) as well as CK2 holoenzyme (K
m 1.1 μM). Different K
m values argue that CK2β(β′) subunit has an inhibitory effect on the activity of both CK2α and CK2α′ towards surviving factor
Svf1. Reconstitution of α′2ββ′ isoform of CK2 holoenzyme shows that β/β′ subunits have regulatory effect depending on the kind of CK2 catalytic subunit.
This effect was not observed in the case of α2ββ′ isoform, which may be due to interaction between Svf1 and regulatory CK2β subunit (shown by co-immunoprecipitation experiments).
Interactions between CK2 subunits and Svf1 protein may have influence on ATP as well as ATP-competitive inhibitors (TBBt and
TBBz) binding. CK2 phosphorylates up to six serine residues in highly acidic peptide K199EVIPESDEEESSADEDDNEDEDEESGDSEEESGSEEESDSEEVEITYED248 of the Svf1 protein in vitro. Presented data may help to elucidate the role of protein kinase CK2 and Svf1 in the regulation
of cell survival pathways. 相似文献
17.
Hiroaki Sawai Takeshi Itoh Kazumi Kokaji Kazuo Shinozuka 《Journal of molecular evolution》1997,45(3):209-215
Oligomerization of α-adenosine 5′-phosphorimidazolide (α-ImpA) has been done in an aqueous solution using a uranyl-ion catalyst
or a poly(U) template as a model process of prebiotic synthesis of RNA with α-glycosidic linkage. α-Oligoriboadenylates up
to hexamer were formed from α-ImpA by the uranyl-ion catalyst. 3′-5′ Linkage was mainly formed in the oligomerization. The
poly(U) template mediated the oligomerization of α-ImpA, but to a very low extent. The yield and chain length of the resulting
α-oligomers were far lower than those of the corresponding β-oligomer formation under the same conditions. Physico-chemical
properties of α-oligoriboadenylates are presented along with those of the corresponding β-oligoriboadenylates. The results
indicate that β-RNA is more advantageous than α-RNA from the points of their synthesis and properties.
Received: 10 February 1997 / Accepted: 31 March 1997 相似文献
18.
Kubiński K Domańska K Sajnaga E Mazur E Zieliński R Szyszka R 《Molecular and cellular biochemistry》2007,295(1-2):229-236
Protein kinase CK2 is a highly conserved Ser/Thr protein kinase that is ubiquitous among eucaryotic organisms and appears
to play an important role in many cellular functions. This enzyme in yeast has a tetrameric structure composed of two catalytic
(α and/or α′) subunits and two regulatory β and β′ subunits. Previously, we have reported isolation from yeast cells four
active forms of CK2, composed of αα′ββ′, α2ββ′, α′2ββ′ and a free α′-catalytic subunit. Now, we report that in Saccharomyces cerevisiae CK2 holoenzyme regulatory β subunit cannot substitute other β′ subunit and only both of them can form fully active enzymatic
unit. We have examined the subunit composition of tetrameric complexes of yeast CK2 by transformation of yeast strains containing
single deletion of the β or β′ regulatory subunits with vectors carrying lacking CKB1 or CKB2 genes. CK2 holoenzyme activity was restored only in cases when both of them were present in the cell. Additional, co-immunoprecypitation
experiments show that polyadenylation factor Fip1 interacts with catalytic α subunits of CK2 and interaction with beta subunits
in the holoenzyme decreases CK2 activity towards this protein substrate. These data may help to elucidate the role of yeast
protein kinase CK2β/β′ subunits in the regulation of holoenzyme assembly and phosphotransferase activity. 相似文献
19.
Lipopolysaccharides (LPSs) of two strains Pragia fontium 97U116 and 27480 were isolated and characterized; they were close to those of other representatives of the family Enterobacteriaceae in fatty acid composition and contained, respectively, 3-hydroxytetradecanoic acid as the predominant component (45.8 and 45.1%), tetradecanoic (23.5 and 28.9%), hexadecanoic (12.6 and 7.9%), hexadecenoic (12.6 and 7.9%), and dodecanoic (4.9 and 4.2%) fatty acids. The O-specific polysaccharides consisted of linear penta- and tetrasaccharide repeating units: →2)-α-D-Galf-(1→3)-α-L-Rhap2Ac-(1→4)-α-D-GlcpNAc-(1→2)-α-L-Rhap-(1→3)-β-D-GlcpNAc-(1→ →4)-β-D-ManpNAc3NAcA-(1→2)-α-L-Rhap-(1→3)-β-L-Rhap-(1→4)-α-D-GlcpNAc-(1→ The LPSs of P. fontium 97U116 and 27480 were serologically active and belonged to different serogroups; they were less toxic than those of strain E. coli O55:B5, but more pyrogenic than the Pyrogenal preparation. 相似文献
20.
Usuki S Ren J Utsunomiya I Cashman NR Inokuchi J Miyatake T 《Neurochemical research》2001,26(4):375-382
We previously reported that ciliary neurotrophic factor (CNTF) increased the serum-free cell survival of immortalized motor neuron-like cells (NSC-34), and addition of the exogenous ganglioside GalNAc4(Neu5Ac3)Gal4GlcCer (GM2) facilitated cell survival together with CNTF. Moreover 1,4 N-acetylgalactosaminyltransferase (GM2 synthase) activity increased in NSC-34 cells cultured with CNTF. We now have examined whether CNTF-induced cell survival is associated with the collaboration between GM2 and the CNTF receptor (CNTF-R). Despite the presence of CNTF (50 ng/ml), anti-CNTF-R antibody caused cell death and prevented the up-regulation of GM2 synthase expression. The addition of GM2 (1 to 20 M) abrogated the anti-CNTF-R antibody effect which shortened cell survival and blocked GM2 synthase activation. Use of [125I]CNTF showed the specificity of CNTF binding in NSC-34 cells in situ. GM2 produced a 5-fold increase in the CNTF binding affinity per cell but did not change the binding site number. The study by metabolic labeling with [1–14C]N-acetyl-D-galactosamine ([14C]GalNAc) showed that biosynthesized GM2 was involved in the immunoprecipitation of CNTF-R. These findings indicate that up-regulated GM2 synthesis induces functional conversion of CNTF-R to the activated state, in which it has affinity for CNTF. We conclude that GM2 is a bio-regulating molecule of CNTF-R in motor neurons. 相似文献