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T Norberg

Publications and source records attributed to T Norberg.

36 records · Page 2Linked to original sources
PubMed1993

The use of human milk fucosyltransferase in the synthesis of tumor-associated trimeric X determinants.

We have studied the fucosylation of a chemically synthesized trimer of N-acetyllactosamine [(LacNAc)3-EtPhNHCOCF3] with a fucosyltransferase preparation from normal human milk, which utilizes both type-1 and type-2 structures, whether sialylated or not. When fucose residues were added enzymically to the (LacNAc)3-EtPhNHCOCF3 hexasaccharide, mono-, di-, or trifucosylated oligosaccharide species were formed, containing the Lewisx determinant (Gal beta 1-->4[Fuc alpha 1-->3]Glc-NAc beta 1-->3). With excess GDP-fucose and prolonged reaction times, the trifucosylated product was formed in almost quantitative yield. Kinetic analysis of the fucosylation reaction indicated that there is a significant difference in the rate of transfer of the first, second and third fucose residues onto the acceptor molecule. The location of the fucose residues in the monofucosylated and difucosylated intermediate products was assessed by analyzing the digests obtained after endo-beta-galactosidase treatment by HPLC and reverse-phase chromatography. In addition, the fucosylated (LacNAc)3-EtPhNHCOCF3 structures were characterized by HPLC and were identified by 400-MHz 1H-NMR spectroscopy. There is a highly preferred order in which the fucosyl residues are attached to (LacN-Ac)3-EtPhNHCOCF3. In the major pathway, the first two fucose residues are transferred with equal preference to the medial (GN3) and proximal (GN1) GlcNAc residues, whereas the third fucose is attached to the distal (GN5) GlcNAc residue. These results are of relevance in understanding the role of alpha-3-fucosyltransferase in the biosynthesis of Lewisx-related cell-surface carbohydrate structures, that function as ligands for selectin-type cell-adhesion molecules and may play a role in the invasion and metastasis of several carcinoma.

Amino Sugars↗
PubMed1993

Quantitative sequence-activity models (QSAM)--tools for sequence design.

Models have been developed that allow the biological activity of a DNA segment to be altered in a desired direction. Partial least squares projections to latent structures (PLS) was used to establish a quantitative model between a numerical description of 68 bp fragments of 25 E.coli promoters and their corresponding quantitative measure of in vivo strength. This quantitative sequence-activity model (QSAM) was used to generate two 68 bp fragments predicted to be more potent promoters than any of those on which the model originally was based. The optimized structures were experimentally verified to be strong promoters in vivo.

Base Composition↗
PubMed1991

Affinity purification of monoclonal antibodies, using a bifunctional oligosaccharide hapten.

A bifunctional hapten was synthesized consisting of a blood group A active tetrasaccharide (A-tetra) and a blood group Le(a) active pentasaccharide lacto-N-fucopentaose II (LNF II), linked to each other with a phenylaminothiourea spacer connecting the reducing ends (A-tetra-LNF II). The hapten was demonstrated to retain both blood group A and Le(a) activity and could be easily bound to both monoclonal anti-A and anti-Le(a) affinity columns. Due to the strong temperature dependence of the two antibodies their binding to oligosaccharides, the bifunctional hapten could be utilized to achieve easy desorption in the final step of affinity purification of either monoclonal anti-Le(a) or anti-A. The system is postulated to have general applicability in affinity purification of any ligate that binds with an avidity too high to achieve non-denaturing desorption.

Antibodies, Monoclonal↗
PubMed1991

Synthesis of the trisaccharide, 2-(p-trifluoroacetamidophenyl)ethyl O-alpha-D-galactopyranosyl-(1-4)-O-beta-D-galactopyranosyl-(1-4)-2-acet amido-2-deoxy-beta-D-glucopyranoside, corresponding to the blood group P1 determinant.

The 2-(p-trifluoroacetamidophenyl)ethyl beta-glycoside of the P1 antigen trisaccharide, O-alpha-D-galactopyranosyl-(1-4)-O-beta-D-galactopyranosyl-(1-4)-2- acetamido-2-deoxy-D-glucopyranose, was synthesized. Thioglycoside intermediates were used as building blocks.

Carbohydrate Sequence↗
PubMed1991

Solid phase synthesis of the fibronectin glycopeptide V(Gal beta 3GalNAc alpha)THPGY, its beta analogue, and the corresponding unglycosylated peptide.

The fibronectin fragment VTHPGY and the corresponding glycopeptides V(Gal beta 3GalNAc alpha)THPGY and V(Gal beta 3GalNAc beta)THPGY were synthesized by the FMOC/solid phase approach. FMOC derivatives of threonine, carrying O-linked, peracetylated Gal beta 3GalNAc chains were used for introduction (HOBt-mediated coupling) of the disaccharide moieties.

Amino Acid Sequence↗
PubMed1991

MCH-induced pigment aggregation in teleost melanophores is associated with a cAMP reduction.

It has previously been shown that alpha 2-adrenoceptors are involved in noradrenaline-induced pigment aggregation within fish melanophores. In the present investigation, melanin concentrating hormone (MCH) elicited pigment aggregation (EC50 approximately 1 x 10(-7) M) that was associated with a significant reduction in the cAMP content; 1 x 10(-7) M MCH reduced the cAMP content from a basal level of 50.4 +/- 2.8 pmol/mg protein to 36.9 +/- 3.8 pmol/mg protein. Like the alpha 2-adrenoceptor-induced pigment aggregation, the MCH response was effectively blocked by the adenylate cyclase stimulator forskolin. These findings suggest that attenuation of cAMP may serve as an intracellular signal transduction mechanism for both MCH and noradrenaline.

Animals↗
PubMed1990

Insulin gene enhancer binding protein Isl-1 is a member of a novel class of proteins containing both a homeo- and a Cys-His domain.

The activity of the rat insulin I gene enhancer is mainly dependent on two cis-acting protein-binding domains. Here we report the isolation of a complementary DNA encoding a protein, Isl-1, that binds to one of these domains. Isl-1 contains a homeodomain with greatest similarity to those of the Caenorhabditis elegans proteins encoded by mec-3 and lin-11. In addition, Isl-1, like the lin-11 and mec-3 gene products, contains a novel Cys-His domain which is reminiscent of known metal-binding regions. Together these proteins define a novel class of proteins containing both a homeo- and a Cys His-domain. Isl-1 is preferentially expressed in cells of pancreatic endocrine origin. If the structural homologies between Isl-1 and the C. elegans gene products reflect functional similarities, a role for Isl-1 in the development of pancreatic endocrine cells could be envisaged.

Amino Acid Sequence↗
PubMed1989

Synthesis of mono- and disaccharide amino-acid derivatives for use in solid phase peptide synthesis.

N-Fluorenylmethyloxycarbonyl-protected serine and threonine derivatives, carrying O-glycosidically alpha- or beta-linked peracetylated beta-D-Galp-(1-3)-D-GalNAcp carbohydrate chains, were prepared. These derivatives are intended for use in solid phase glycopeptide synthesis. Suitably protected mono- and disaccharide thioglycosides were used as carbohydrate intermediates. These were activated by treatment with bromine to give the glycosyl bromides, which were then used in silver triflate-promoted glycosidations of N-fluorenylmethyloxycarbonyl amino-acid phenacyl esters. Removal of the phenacyl esters with zinc gave the target free acids.

Amino Acids↗
PubMed1989

Synthesis of an H type 2 and a Y (Le(y)) glycoside from thioglycoside intermediates.

The trisaccharide 2-(p-trifluoroacetamidophenyl)ethyl 2-acetamido-2- deoxy-4-O-[2-O-(alpha-L-fucopyranosyl)-beta-D-galactopyranosyl]-beta-D- glucopyranoside 1 and the tetrasaccharide 2-(p-trifluoroacetamidophenyl)ethyl 2-acetamido-2-deoxy-3-O-(alpha-L- fucopyranosyl)-4-O-[2-O-(alpha-L-fucopyranosyl)-beta-D-galactopyranosyl]- beta-D-glucopyranoside 2 were synthesized. Thioglycosides, suitably protected, activated directly with methyl trifluoromethanesulfonate or dimethyl(methylthio)sulfonium tetrafluoroborate or activated after bromine treatment with halophilic reagents, were used as glycosyl donors in the construction of the glycosidic linkages.

Carbohydrate Conformation↗
PubMed1988

Synthesis of a dimeric Lewis X hexasaccharide derivative corresponding to a tumor-associated glycolipid.

The dimeric Lewis X hexasaccharide p-trifluoroacetamidophenylethyl O-beta-D-galactopyranosyl-(1----4)-O-[alpha-L-fucopyranosyl-(1----3)]-O- (2- acetamido-2-deoxy-beta-D-glucopyranosyl)-(1----3)-O-beta-D-galactopyrano syl- (1----4)-O-[alpha-L-fucopyranosyl-(1----3)]-2-acetamido-2-deoxy-beta-D- glucopyranoside (14), which is a derivative of a tumor-associated glycolipid, was synthesized from thioglycoside intermediates. A protected disaccharide was used as a key-intermediate for synthesis of the p-nitrophenylethyl glycoside of suitably protected O-beta-D-Galp-(1----4)-O-beta-D-GlcpN-(1----3)-O-beta-D-Galp-(1--- -4)-beta-D- GlcpN, which, after selective deblocking, was di-L-fucosylated and deprotected to give 14.

Biomarkers, Tumor↗
PubMed1985

Synthesis of the methyl and 1-octyl glycosides of the P-antigen tetrasaccharide (globotetraose).

The methyl and 1-octyl beta-glycosides of the P-antigen tetrasaccharide [globotetraose, beta-D-GalpNAc-(1----3)-alpha-D-Galp-(1----4)-beta-D-Galp-(1----4) -D-Glc] were synthesised from a tetrasaccharide precursor, prepared using methyl disaccharide 1-thioglycosides as intermediates. In the key glycosidation with silver triflate, HO-2 was used as an alpha-directing group in the glycosyl bromide.

Carbohydrate Conformation↗
PubMed1984

The conformation of Salmonella O-antigenic polysaccharide chains of serogroups A, B, and D1 predicted by semi-empirical, Hard-Sphere (HSEA) calculations.

The preferred conformation of the tetrasaccharide repeating units of Salmonella Serogroups A, B, and D1 have been calculated. The semiempirical calculations used the Hard-Sphere, Exo-Anomeric (HSEA) approach to derive a conformational model, which could be used to assist the interpretation of conformations significantly populated in aqueous solution. The calculated model was extended to include a pentasaccharide repeating unit bearing an alpha-D-glucopyranose-branch point. The 3,6-dideoxyhexose to D-mannose linkage was shown to possess a steep energy surface with a minimum, which results in good exposure of the dideoxyhexose O-2 and O-4 atoms. Stereochemical changes involving the equatorial or axial disposition of these atoms are the distinguishing structural features of the A, B, and D1 serogroups. A lipophilic surface involving the 6-deoxy groups of the dideoxy-D-hexose and L-rhamnose residues was identified, and the possible implications of these features in antigenic determinants is discussed. The preferred conformation predicted by the HSEA method correlates with the known antigenic specificities of polysaccharides belonging to these Salmonella serogroups.

Carbohydrate Conformation↗
PubMed1984

The conformation of Salmonella O-antigenic oligosaccharides of serogroups A, B, and D1 inferred from 1H- and 13C-nuclear magnetic resonance spectroscopy.

The conformational model derived by the HSEA calculation method was used to interpret the n.m.r. data for solutions of oligosaccharides corresponding to the Salmonella serogroups A, B, and D1 antigenic determinants. The favored conformer, derived by calculation, accounted for the observed, chemical-shift changes and accurately predicted the existence and magnitude of inter-ring proton n.O.e.'s. Extensive proton-density and compression of proton, Van der Waals radii were correlated with deshielding of specific proton-resonances. The model of lipopolysaccharide conformation accounts for the known antigenic properties of Salmonella O-antigens.

Carbohydrate Conformation↗
PubMed1984

Specific identification of Salmonella serogroup E antigen O3 by immunofluorescence and coagglutination with antiserum elicited by a synthetic trisaccharide-bovine serum albumin glycoconjugate.

Antiserum specific for Salmonella O3 antigen was raised by immunization of rabbits with an artificial glycoconjugate consisting of the synthetic trisaccharide beta-D- Manp (1----4)-alpha-L- Rhap (1----3)-alpha-D-Galp covalently linked to bovine serum albumin (beta- MRG -BSA). Enzyme immunoassays showed that only lipopolysaccharides extracted from Salmonella serogroup E (O3 antigen-containing) bacteria bound the antiserum. The usefulness of the beta- MRG -BSA antiserum for rapid and accurate identification of Salmonella isolates of serogroup E was shown by indirect immunofluorescence tests in which 50 Salmonella strains of serogroup E 1-4 were correctly identified from among 651 intestinal strains investigated. The finding that one strain of Citrobacter freundii was positive in immunofluorescence tests with this antiserum is readily explained by the known cross-reactivity between some C. freundii strains and Salmonella spp. strains of serogroup E. As expected, the specificity of the antiserum for the O3 antigen could further be demonstrated in coagglutination tests with staphylococci sensitized with beta- MRG -BSA antiserum.

Agglutination Tests↗
PubMed1982

Synthesis of type 2 human blood-group antigenic determinants. The H, X, and Y haptens and variations of the H type 2 determinant as probes for the combining site of the lectin I of Ulex europaeus.

Chemical syntheses of the human blood-group antigenic determinants derived from N-acetyllactosamine are described; namely, the 6-deoxy derivative, the 4'-epimer, and the 5 H type 2 [alpha LFuc(1 to 2)beta DGal-(1 to 4)beta DGlcNAc], X [beta DGal(1 to 4)[alpha LFuc(1 to 3)[beta DGlcNAc], and Y [alpha LFuc-(1 to 2)beta DGal(1 to 4)[alpha LFuc(1 to 3)]beta DGlcNAc] determinants as glycosides of 8-carboxymethyloctanol. In order to study the binding of the H type 2 determinant with the lectin I of Ulex europaeus, structures designed to specifically alter the hydrophilic and hydrophobic portions of the H type 2 determinant were also prepared; namely, the 6-deoxy derivative, the 4'-epimer, and the 5"-nor-homolog. The use of these structures, together with the H type 1 hapten and the N-deacetylated forms of both the H type 1 and H type 2 determinants, as inhibitors of the agglutination of O red cells by the lectin allowed the conclusion that the binding of the H type 2 determinant is hydrophobic; the binding involves a wedge-like portion of the determinant that is basically hydrophobic, except for the 5-hydroxymethyl group, which is at the tip of the wedge and forms an intramolecular hydrogen-bond with O-5 for acceptance by a hydrophobic cleft at the surface of the lectin. Blocking procedures involving alkoxymethyl groups and new experiences involving glycosylation reactions are described.

Carbohydrate Conformation↗
PubMed1982

A 500-MHz proton-magnetic-resonance study of several fragments of the carbohydrate-protein linkage region commonly occurring in proteoglycans.

The proton-magnetic-resonance spectra of three partial structures of the carbohydrate-protein linkage region that frequently occurs in proteoglycans, namely, beta-D-Galp-(1 leads to 3)-beta-D-Galp-(1 leads to 4)-beta-D-Xylp-(1 leads to O)-L-Ser, were recorded in 2H2O at 500 MHz; they could be completely interpreted, both for the glyco-serines and for the corresponding glyco-xylitols. The chemical shifts and the coupling constants were refined by computer simulation of the spectra. The change in the chemical shift of H-4 of a D-galactopyranosyl residue upon substitution at C-3 by a beta-D-galactopyranosyl group is proposed to be characteristic for this particular attachment, making H-4 of galactose a structural-reporter group. The three constituting monosaccharides adopt the 4C1 (D) ring conformation. The terminal galactopyranosyl group and the internal galactopyranosyl residue differ as to the population of rotamers around the C-5/C-6 axis. Concomitantly, the flexibility of their glycosidic linkages is distinct.

Carbohydrates↗