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Biomedical subjects

F Inagaki

Publications and source records attributed to F Inagaki.

At least 127 records · Page 7Linked to original sources
PubMed1988

Immunoaffinity isolation of a sialyl-Le(a) oligosaccharide from human milk.

A cancer-associated antigen, sialyl-Le(a) oligosaccharide, was isolated from human milk using a monoclonal antibody recognizing carbohydrate moieties of mucin-type glycoproteins. The structure was identified as: (Formula: see text) based on 500-MHz 1H-NMR spectroscopy. This oligosaccharide comprises 0.07% of sialyloligosaccharides in human milk. The NMR spectra of two fellow oligosaccharides, Le(a) oligosaccharide (or lacto-N-fucopentaose II) and LS-tetrasaccharide a, are also given.

Antibodies, Monoclonal↗
PubMed1988

Melibiosylceramide as the sole ceramide dihexoside from the eggs of the sea urchin, Anthocidaris crassispina.

Melibiosylceramide (Gal alpha 1-6Glc beta 1-1Cer) was found as the sole ceramide dihexoside from the eggs of the sea urchin, Anthocidaris crassispina. Ceramide monohexoside of the eggs consisted only of glucosylceramide (Glc beta 1-1Cer). These lipids were purified by successive column chromatographies on DEAE-Sephadex A-25, silicic acid and Florisil, and identified by gas-liquid chromatography, negative-ion fast atom bombardment mass spectrometry and proton nuclear magnetic resonance spectroscopy as well as methylation analysis. Long-chain base compositions of both lipids were almost identical and comprised n-C18-phytosphingosine and small amounts of its homologs (C17-C19). Fatty acid compositions were qualitatively very similar, but the glucosylceramide contained more 2-hydroxy fatty acid than the melibiosylceramide. Although the chain length of fatty acids was distributed over a wide range, six major fatty acids, namely 22:1, 23:1, 24:1, 22h:1, 23h:1 and 24h:1, constituted more than 92% of the fatty acid content in these lipids.

Animals↗
PubMed1988

A comparative 1H NMR study of mouse alpha(1-53) and beta(2-53) epidermal growth factors.

The three-dimensional structure of the mouse epidermal growth factor (EGF) in solution was studied by comparison of the 1H NMR spectra of alpha EGF (1-53) and beta EGF (2-53, des-asparaginyl 1 form). Using pH dependence of chemical shifts and a two-dimensional difference spectrum, the effect of the N-terminal deletion was investigated based on the complete assignment of the proton resonances. The affected residues were all found to be located exactly in the triple-stranded, beta-sheet core in the N-terminal domain of the EGF molecule.

Amino Acids↗
PubMed1987

Characterization of tumor-associated fucogangliosides from PC 12 pheochromocytoma cells.

PC 12h pheochromocytoma cells were subcutaneously transplanted into rat. We found the transplanted tumors accumulated some fucogangliosides associated with PC 12 cells. These gangliosides were isolated and purified by DEAE-Sephadex A-25 and Iatrobeads column chromatographies. Their structures were determined by fast atom bombardment mass spectrometry, proton nuclear magnetic resonance spectrometry, permethylation study, and sequential degradation using various exoglycosidases and mild acid hydrolysis. Two tumor-associated fucogangliosides were found to possess the blood group B determinant as follows: G6: IV2Fuc alpha, IV3Gal alpha, II3NeuAc, GgOse4Cer; G11: IV2Fuc alpha, IV3Gal alpha, II3 (NeuAc)2, GgOse4Cer. A ganglioside with the similar structure as ganglioside G6 was isolated from rat hepatoma cells (Holmes, E.H., and Hakomori, S-I. (1982) J. Biol. Chem. 257, 7698-7703). However, ganglioside G11 has not previously been reported in the literature. These fucogangliosides reacted with the monoclonal antibody prepared by immunizing mice with PC 12h cells. Other fucogangliosides were also found to accumulate in the transplanted tumor tissues. They were identified as fucosyl-GM1 and fucosyl-GDlb. These fucogangliosides did not react with the monoclonal antibody against PC 12h cells.

Adrenal Gland Neoplasms↗
PubMed1987

Analysis of NMR spectra of sugar chains of glycolipids by 1D homonuclear Hartmann-Hahn and NOE experiments.

We applied 1D homonuclear Hartmann-Hahn (1D-HOHAHA) and difference NOE experiments to determine the chemical structure of Forssman's antigen, a glycolipid purified from sheep red blood cells. The subspectra corresponding to the individual sugar components were extracted from overlapping proton resonances by selective excitation of the anomeric proton resonances, so that unambiguous assignments of the sugar proton resonances were accomplished. Then, difference NOE experiments were performed to determine the linkage of the sugar units. The present procedure was found to be useful for the structure determination of glycoconjugates and also reduces the amount of samples and machine time.

Animals↗
PubMed1987

Analysis of NMR spectra of sugar chains of glycolipids by multiple relayed COSY and 2D homonuclear Hartman-Hahn spectroscopy.

We applied multiple relayed COSY and 2D homonuclear Hartman-Hahn spectroscopy to globoside, a glycolipid purified from human red blood cells. The subspectra corresponding to individual sugar components were extracted even from overlapping proton resonances by taking the cross sections of 2D spectra parallel to the F2 axis at anomeric proton resonances, so that unambiguous assignments of sugar proton resonances were accomplished.

Galactosamine↗
PubMed1987

A new extended globoglycolipid carrying the stage specific embryonic antigen-1 (SSEA-1) determinant in mouse kidney.

Two neutral glycolipids carrying the stage specific embryonic antigen-1 (SSEA-1) and SSEA-3 determinants, respectively, were purified from mouse kidney by a combination of column chromatographies and droplet counter-current chromatography. The structures of the glycolipids (GL-X and GL-Y) were determined by means of GLC, 1H-NMR spectroscopy, negative-ion fast atom bombardment mass spectrometry, a methylation study, and sequential degradation. GL-X was demonstrated to be galactosyl beta 1-3globotetraosylceramide, the structure of which had already been characterized to be that of SSEA-3 by Kannagi et al. [1983) J. Biol. Chem. 258, 8934-8942). GL-Y was a new glycolipid containing fucose, galactose, glucose, N-acetylgalactosamine, and N-acetylglucosamine in a molar ratio of 1:4:1:1:1. The methylation study results indicated that it contained 3 mol of terminal sugars composed of 2 mol of galactose and 1 mol of fucose with two branching points at N-acetylgalactosamine and N-acetylglucosamine. From the data obtained by 1H-NMR spectroscopy, mass spectrometry, and a binding assay using an anti-SSEA-1 monoclonal antibody (PM81) cloned by Ball et al. [1983) J. Immunol. 130, 2937-2941), we propose the structure of GL-Y to be Gal beta 1-4GlcNAc beta 1-6GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc beta 1-ceramide. (sequence; see text) Fuc alpha 1 Gal beta 1 This is the first report on the isolation and characterization of a glycolipid carrying the SSEA-1 determinant on its globo-core structure.

Animals↗
PubMed1987

A new ganglioside showing choleragenoid-binding activity in mouse spleen.

A new ganglioside showing choleragenoid-binding activity was purified from mouse spleen and characterized. From the results of sugar-composition analysis, enzymatic hydrolysis, a permethylation study, 1H-NMR spectroscopy, and negative-ion fast atom bombardment mass spectrometry, the structure of the ganglioside was determined to be as follows: Gal beta 1-3GalNAc beta 1-4Gal beta 1-3GalNAc beta 1-4Gal beta 1-4Glc beta 1-1'ceramide 3----NeuGc alpha 2 This ganglioside contains a terminal tetrasaccharide structure identical with that of II3NeuGc alpha-Gg4Cer (GM1(NeuGc]. By means of a TLC-immunobinding assay and an enzyme-linked immunosorbent assay, the ganglioside was demonstrated to have almost the same choleragenoid-binding activity as GM1. Another ganglioside, that migrated faster than the new choleragenoid-binding ganglioside, was also purified from the same source material and identified as IV4GalNAc beta,IV3NeuGc alpha-Gg4Cer (GalNAc-GM1b(NeuGc]. Since, in the previous study, we demonstrated the existence of IV3NeuGc alpha-Gg4Cer (GM1b(NeuGc] in mouse spleen (Nakamura, K. et al. (1984) J. Biochem. 96, 949-957), the results of this study suggest that the new choleragenoid-binding ganglioside is synthesized from GM1b(NeuGc) through GalNAc-GM1b(NeuGc).

Animals↗
PubMed1986

Conformations of fibroblast and E. coli-derived recombinant human interferon-beta s as studied by nuclear magnetic resonance and circular dichroism.

The conformations of fibroblast and E. coli-derived recombinant human interferon-beta s were studied by circular dichroism and nuclear magnetic resonance spectroscopy in the acidic pH region of 4.6 to 1.6. Both interferons have very similar conformations with high alpha-helix contents (approximately 70%). These results suggest that glycosylation does not appreciably change the conformation of human interferon-beta. Moreover, a slow conformational change is observed below pH 2.0, which induces the disruption of beta-sheets.

Circular Dichroism↗
PubMed1986

Newly developed circulating blood volume-monitoring system and its clinical application for measuring changes in blood volume during hemofiltration.

Blood and ultrafiltrate electrical resistivities were continuously monitored during hemofiltration. By substituting these values into a previously developed equation that was modified by the authors, the hematocrit value was determined; and blood volume change was obtained from the change in this value. The following facts were discovered as the result of monitoring the blood volume change during treatment: When the body position was changed from the reclining to the sitting position, the blood volume decreased by 4.2 +/- 0.3%. With the body water removal rate kept constant, the body position recumbent, and the subject fasted, the blood volume gradually decreased. However, when the position was changed from reclining to sitting for food intake and once more returned to the supine position after the meal, the blood volume was greatly affected by the change in position and the food intake.

Blood↗
PubMed1985

Molecular conformation of alpha-bungarotoxin as studied by nuclear magnetic resonance and circular dichroism.

We have examined the circular dichroism and nuclear magnetic resonance spectra of a long neurotoxin, alpha-bungarotoxin, over a wide range of pH values and temperatures, and under high salt conditions. The observations are interpreted partly in terms of the known crystal structure of this polypeptide. We support earlier findings of a greater degree of beta-sheet structure in solution than has been reported by X-ray crystallography and, importantly, the invariant residue associated with neurotoxicity, Trp29, is shown to be in a similar environment to that found in alpha-cobratoxin and LS III from Laticauda semifasciata. The implications of this observation for structure/function relationships are outlined.

Amino Acid Sequence↗
PubMed1985

Binding modes of inhibitors to ribonuclease T1 as studied by nuclear magnetic resonance.

The binding modes of inhibitors to ribonuclease T1 (RNase T1) were studied by the analyses of 270-MHz proton NMR spectra. The chemical shift changes upon binding of phosphate, guanosine, 2'-GMP, 3'-GMP, 5'-GMP, and guanosine 3',5'-bis(phosphate) were observed as high field shifted methyl proton resonances of RNase T1. One methyl resonance was shifted upon binding of phosphate and guanosine nucleotides but not upon binding of guanosine. Four other methyl resonances were shifted upon binding of guanosine and guanosine nucleotides but not upon binding of phosphate. From the analyses of nuclear Overhauser effects for the pair of H8 and H1' protons, together with the vicinal coupling constants for the pair of H1' and H2' protons, the conformation of the guanosine moiety as bound to RNase T1 is found to be C3'-endo-syn for 2'-GMP and 3'-GMP and C3'-endo-anti for 5'-GMP and guanosine 3',5'-bis(phosphate). These observations suggest that RNase T1 probably has specific binding sites for the guanine base and 3'-phosphate group (P1 site) but not for the 5'-phosphate group (PO site) or the ribose ring. The weak binding of guanosine 3',5'-bis(phosphate) and 5'-GMP to RNase T1 is achieved by taking the anti form about the glycosyl bond. The productive binding to RNase T1 probably requires the syn form of the guanosine moiety of RNA substrates.

Endoribonucleases↗