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S Tsuji

Publications and source records attributed to S Tsuji.

At least 19 recordsLinked to original sources
PubMed1995

A developmentally regulated member of the sialyltransferase family (ST8Sia II, STX) is a polysialic acid synthase.

We found polysialic acid synthase activity of ST8Sia II (STX) in vitro and in vivo. Previously, we showed that mouse ST8Sia II exhibits alpha 2,3-sialylated N-glycan alpha 2,8-sialyltransferase activity, but the polysialic acid synthase activity of ST8Sia II was not detected at that time [Kojima, N. et al. (1995) FEBS Lett. 360, 1-4]. When fetuin was [14C]sialylated with ST8Sia II and then its N-linked oligosaccharides were analyzed, a part of the N-linked oligosaccharides was eluted in the void volume from a Sephadex G-50 column, and was eluted in the void volume from a Sephadex G-50 column, and was eluted from the DEAE-Toyopearl column at almost the same salt concentration as that where colomic acid was eluted. In addition, a series of 14C-labeled oligo-sialic acids were obtained from the oligosaccharides on partial mild acid hydrolysis. These results indicated that a part of N-linked oligosaccharides of fetuin were polysialylated with ST8Sia II. Transfection of ST8Sia II gene into several cell lines including NIH3T3 led to the expression of polysialic acids on the cell surface. Thus, ST8Sia II can directly synthesize polysialic acid chains on alpha 2,3-sialylated N-linked oligosaccharides of glycoproteins without any initiator sialytransferase.

3T3 Cells↗
PubMed1995

GTP cyclohydrolase I gene in hereditary progressive dystonia with marked diurnal fluctuation.

We previously reported four different mutations in the coding region of GTP cyclohydrolase I (GCH-I) gene in patients with hereditary progressive dystonia with marked diurnal fluctuation (HPD). We found two independent new mutations (leucine 79 proline and a deletion in exon 4) in patients with HPD. We also found four families of HPD without any mutations in the coding region of GCH-I gene.

Base Sequence↗
PubMed1995

Direct detection of expanded (CAG/CTG) repeats in the myotonin-protein kinase genes of myotonic dystrophy patients using a high-stringency hybridization method.

Recent discoveries of genes containing (CAG/CTG) repeats as the causative genes for neuromuscular diseases suggest that trinucleotide repeat expansions are associated with a number of other neurodegenerative diseases, particularly in those showing genetic anticipation. In order to efficiently identify expanded trinucleotide repeats, we have developed a novel method using a single-stranded probe containing (CAG)55, which allows direct detection of expanded (CAG/CTG) repeats in the myotonin-protein kinase (Mt-PK) genes of patients with myotonic dystrophy without the need for any prior information of chromosomal localizations of the disease genes. This method is expected to be useful for identifying novel genes for diseases associated with expanded (CAG/CTG) repeats.

Base Sequence↗
PubMed1995

Molecular cloning of Sia alpha 2,3Gal beta 1,4GlcNAc alpha 2,8-sialyltransferase from mouse brain.

A cDNA encoding a new alpha 2,8-sialyltransferase (ST8Sia III), which exhibits activity toward the Sia alpha 2,3Gal beta 1, 4GlcNAc sequences of N-linked oligosaccharides, was cloned from mouse brain by means of the polymerase chain reaction-based approach. The predicted amino acid sequence of ST8Sia III showed 27.6 and 34.4% identity with those of so far cloned mouse alpha 2,8-sialyltransferases, i.e. GD3 synthase (ST8Sia I) and STX (ST8Sia II), respectively. Transfection of the protein A-fused ST8Sia III gene into COS-7 cells led to alpha 2,8-sialyltransferase activity toward sialylated glycoproteins and alpha 2,3-sialylated glycosphingolipids, such as alpha 2,3-sialylparagloboside and GM3. However, the kinetic properties of ST8Sia III revealed that it is much more specific to N-linked oligosaccharides of glycoproteins than glycosphingolipids. The expression pattern of the ST8Sia III gene was clearly different from those of other alpha 2,8-sialyltransferase genes. The expression of the ST8Sia III gene was tissue and stage specific. The ST8Sia III gene was expressed only in brain and testis, and it appeared first in 20 postcoitum embryonal brain and then decreased. Therefore, the new alpha 2,8-sialyltransferase is closely involved in brain development.

Amino Acid Sequence↗
PubMed1995

Characterization of the phosphatidylserine-binding region of rat MARCKS (myristoylated, alanine-rich protein kinase C substrate). Its regulation through phosphorylation of serine 152.

We reported previously that recombinant myristoylated, alanine-rich protein kinase C substrate (MARCKS) expressed in Escherichia coli as well as MARCKS purified from rat brain specifically bound to phosphatidylserine (PS) in a calcium-independent manner and that the binding was regulated through phosphorylation of MARCKS (Nakaoka, T., Kojima, N., Hamamoto, T., Kurosawa, N., Lee, Y. C., Kawasaki, H., Suzuki, K., and Tsuji, S. (1993) J. Biochem. (Tokyo) 114, 449-452). In this study, to identify the minimum PS-binding region of MARCKS and the regulatory phosphorylation site, the binding of MARCKS to PS was examined in deletion mutants producing glutathione S-transferase (GST) fusion proteins. The mutant proteins GST-6-180 and GST-127-160 had almost the same ability to bind to immobilized PS as MARCKS purified from rat brain, whereas GST-127-152 did not bind to it. In addition, the binding of GST-6-156 to immobilized PS was 62% of that of GST-6-180, but that of GST-6-152 was only 8% and that of GST-6-135 was not detected. The effect of phosphorylation by protein kinase C was examined in several mutants of GST-6-180 whose serine residues were substituted with alanine. After phosphorylation, the mutants GST-6-180[S156A and S163A], GST-6-180]S156A], and GST-6-180[S163A] did not bind to immobilized PS like native MARCKS and GST-6-180. However, even after phosphorylation, GST-6-180-[S152A] and GST-6-180[S152A and S156A] could bind to immobilized PS. These results strongly suggest that MARCKS binds to PS molecules in the inner leaflet of the plasma membrane through residues 127-156, with residues 153-156 (FKKS) being particularly important in the binding of MARCKS to PS, and that the binding is regulated through the protein kinase C-catalyzed phosphorylation of the serine at residue 152.

Amino Acid Sequence↗
PubMed1995

Molecular cloning and expression of chick Gal beta 1,3GalNAc alpha 2,3-sialyltransferase.

A cDNA clone encoding chick Gal beta 1,3GalNAc alpha 2,3-sialyltransferase (ST3Gal I) was isolated from a chick embryo brain cDNA library. The cDNA sequence included an open reading frame coding for 342 amino acids, and the deduced amino acid sequence showed 64% identity with that of the mouse enzyme. Northern blot analysis of chick embryos revealed that the ST3Gal I gene was expressed in early embryonic stages. The identity of the enzyme was confirmed by construction of a recombinant sialyltransferase in which the N-terminal part including the cytoplasmic tail and signal anchor domain was replaced with an immunoglobulin signal peptide sequence. This enzyme expressed in COS-7 cells exhibited transferase activity similar to that of mouse ST3Gal I.

Amino Acid Sequence↗
PubMed1995

Donor substrate specificities of Gal beta 1,4GlcNAc alpha 2,6-sialyltransferase and Gal beta 1,3GalNAc alpha 2,3-sialyltransferase: comparison of N-acetyl and N-glycolylneuraminic acids.

Using cloned sialyltransferases, Gal beta 1,3GalNAc alpha 2,3-sialyltransferase (ST3Gal I) and Gal beta 1,4GlcNAc alpha 2,6-sialyltransferase (ST6Gal I) from both chicken and mouse, CMP-NeuAc and CMP-NeuGc were compared as donor substrates with pyridylamino-oligo-saccharides as acceptors. ST6Gal I showed 4-7-times higher activity toward CMP-NeuGc than CMP-NeuAc, while for ST3Gal I there was no significant difference between them, irrespective of the origin of the enzymes. Also, the difference in donor substrate (i.e., NeuAc and NeuGc) had little effect on the preference to acceptor substrates of these enzymes. Thus, the results showed that the cloned sialyltransferases can utilize both CMP-NeuAc and CMP-NeuGc as donor substrates, and that the preference difference between the sialyltransferases to CMP-NeuGc and CMP-NeuAc could, at least partly, explain the discrepancy in the ratio of NeuAc and NeuGc in glycolipids and glycoproteins in individual tissues.

Animals↗
PubMed1995

Molecular cloning and expression of two types of rabbit beta-galactoside alpha 1,2-fucosyltransferase.

Two DNA clones encoding rabbit beta-galactoside alpha 1,2-fucosyltransferase (RFT-I and RFT-II) have been isolated from a rabbit genomic DNA library. The DNA sequences revealed open reading frames coding for 373 (RFT-I) and 354 (RFT-II) amino acids, respectively. The deduced amino acid sequences of RFT-I and RFT-II showed 56% identity with each other, and that of RFT-I showed 80% identity with that of human H blood type alpha 1,2-fucosyltransferase. Northern blot analysis of embryo and adult rabbit tissues revealed that the RFT-I gene was expressed in adult brain, and that the RFT-II gene was expressed in salivary and lactating mammary glands. The identities of these enzymes were confirmed by constructing recombinant fucosyltransferases in which the N-terminal part including the cytoplasmic tail and signal anchor domain was replaced with the immunoglobulin signal peptide sequence. RFT-I expressed in COS-7 cells exhibited similar transferase activity to that of human H blood type alpha 1,2-fucosyltransferase. RFT-II expressed in COS-7 cells showed higher affinity for type 1 (Gal beta 1,3GlcNAc) and type 3 (Gal beta 1,3GalNAc) acceptors than type 2 (Gal beta 1,4GlcNAc) ones, which suggested that RFT-II was a putative secretor-type alpha 1,2-fucosyltransferase.

Amino Acid Sequence↗
PubMed1995

Enzymatic activity of a developmentally regulated member of the sialyltransferase family (STX): evidence for alpha 2,8-sialyltransferase activity toward N-linked oligosaccharides.

We have detected sialyltransferase activity of recombinant mouse STX, which was cloned from rat brain as a new member of the sialyltransferase family, but sialyltransferase activity of which had not been detected previously [Livingston and Paulson, J. Biol. Chem. (1993) 268, 11504-11507]. The activity of mouse STX was specific toward sialylated glycoproteins. N-Glycanase treatment and linkage-specific sialidase treatment of glycoproteins revealed that STX transfers sialic acids through alpha 2,8-linkages to only N-linked oligosaccharides of glycoproteins. However, polymerase activity for polysialic acid synthesis was not detected for this sialyltransferase. Since this alpha 2,8-sialyltransferase gene is highly restricted in fetal and newborn brain, it may be involved in the polysialylation of glycoproteins, especially of N-CAM.

Animals↗
PubMed1995

Lactate rise in the basal ganglia accompanying finger movements: a localized 1H-MRS study.

To investigate whether motor activation can cause lactate elevation, we observed the metabolic changes in seven right-handed volunteers by localized 1H-magnetic resonance spectroscopy (MRS). The volume of interest (VOI) was centered on a region including portions of the putamen and globus pallidus. Finger opposition movements were applied as the motor tasks. On the side contralateral to the finger movements, lactate rose in all subjects. No lactate rise occurred in basal ganglia ipsilateral to the movements.

Basal Ganglia↗
PubMed1995

Plasticity of nerve afferents to nigrostriatal neurons in Parkinson's disease.

Clinical symptoms in Parkinson's disease do not appear until almost total depletion of dopamine has occurred in the striatum, suggesting the existence of compensatory mechanisms to offset the loss of nigrostriatal dopaminergic neurons. This compensation has been attributed mainly to an increased turnover of dopamine in the remaining dopaminergic neurons. Besides this biochemical phenomenon intrinsic to dopaminergic neurons, we tested whether morphological changes in the nerve afferents to the dopaminergic neurons could participate in these compensatory mechanisms. The afferents to the dendrites of dopaminergic neurons were analyzed ultrastructurally in the substantia nigra of parkinsonian patients and matched controls, using simultaneous histochemical detection of acetylcholine-like cation and tyrosine hydroxylase. The size of acetylcholine-like cation-containing terminals in contact with dopaminergic dendrites increased significantly by 38% in the substantia nigra of parkinsonian patients; whereas their number per section of dopaminergic dendrite showed an increase of 60%, although not reaching statistical significance. The number of the terminals devoid of acetylcholine-like cation per section of dopaminergic dendrite decreased significantly by 52% in the substantia nigra of parkinsonian patients. These results suggest (1) a plasticity of excitatory cholinergic neurons targeting nigral dopaminergic neurons and (2) an involution of noncholinergic nerve terminals, mostly originating from inhibitory nigral, pallidal, and striatal GABAergic neurons. The findings provide evidence of a capacity for neuronal plasticity in the elderly human brain, even in the presence of neurodegenerative disorders.

Acetylcholine↗
PubMed1995

The gene for hereditary progressive dystonia with marked diurnal fluctuation maps to chromosome 14q.

Hereditary progressive dystonia with marked diurnal fluctuation (HPD) is a childhood-onset, postural dystonia that is characterized by marked diurnal fluctuation and a dramatic response to levodopa. Recently, the gene for dopa-responsive dystonia (DRD), an autosomal dominant dystonia showing similarly marked response to levodopa, has been mapped to chromosome 14q. Since HPD and DRD share many clinical characteristics, we have analyzed microsatellite polymorphisms in the region of the DRD locus and obtained a maximal lod score of 2.0 at D14S52 without obligate recombination events in the affected individuals. The results strongly suggest that HPD and DRD are to be caused by mutations in the same gene on the long arm of chromosome 14.

Chromosome Mapping↗
PubMed1995

Quantitation of heteroplasmy of mitochondrial tRNA(Leu(UUR)) gene using PCR-SSCP.

We have devised a novel method for quantitative analysis of the MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes) tRNA(Leu(UUR)) mutation of mitochondrial DNA using a PCR-SSCP (polymerase chain reaction-single-strand conformation polymorphism) method, and compared the results obtained using the PCR-SSCP method with those obtained using other methods including Southern blotting, last one cycle hot PCR, and conventional PCR-RFLP (restriction fragment length polymorphism). The standard curve obtained using the PCR-SSCP method is linear, with a correlation coefficient of 0.999; it was determined that this method is more accurate than other methods for quantitative analysis. The PCR-SSCP method does not require restriction digestions, thereby avoiding potential problems of partial digestions or heteroduplex formation during PCR. The method is quite simple and should have a broad range of application for quantitation of mutant mtDNAs in various mitochondrial encephalomyopathies. We applied the method for quantitation of mutant mitochondrial DNA carrying a single base substitution in the tRNA(Leu(UUR)) gene in two autopsied cases of MELAS. In both cases, the mutant mtDNA is abundantly present (82-95%) withd little variation among tissues.

Base Sequence↗
PubMed1995

A quantitative approach to technetium-99m ethyl cysteinate dimer: a comparison with technetium-99m hexamethylpropylene amine oxime.

To develop non-invasive regional cerebral blood flow (rCBF) measurements using technetium-99m ethyl cysteinate dimer (99mTc-ECD) and single-photon emission tomography (SPET), the same graphical analysis as was described in our previous reports using technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO) was applied to time-activity data for the aortic arch and brain hemispheres after intravenous injection of 99mTc-ECD. Hemispherical brain perfusion indices (BPI) for 99mTc-ECD showed a highly significant correlation (n = 22, r = 0.935, P = 0.0001) with those for 99mTc-HMPAO in 11 patients who underwent both tracer studies. Using both linear regression line equations between 99mTc-ECD BPI and 99mTc-HMPAO BPI and between 99mTc-HMPAO BPI and mean cerebral blood flow (CBF) values obtained from a xenon-133 inhalation SPET method in a previous study, 99mTc-ECD BPI was converted to 133Xe CBF values (y = 2.60 chi + 9.8). Then raw SPET images of 99mTc-ECD were converted to rCBF maps using Lassen's correction algorithm. In this algorithm, the correction factor alpha was fixed to 1.5, 2.6 and infinite. In the comparison of rCBF values for 99mTc-ECD SPET with those for 99mTc-HMPAO SPET in 396 regions of interest in the aforementioned 11 patients, the fixed correction factor alpha of 2.6 gave nearly the same rCBF values for 99mTc-ECD (50.1 +/- 16.9 ml/100 g/min, mean +/- SD) as for 99mTc-HMPAO (49.9 +/- 17.3 ml/100 g/min). In conclusion, the same non-invasive method as has been used in 99mTc-HMPAO studies is applicable to a 99mTc-ECD study for the measurement of rCBF without any blood sampling.

Adult↗
PubMed1995

Roles of hepatocyte growth factor and its receptor in gastric mucosa. A cell biological and molecular biological study.

Hepatocyte growth factor (HGF) stimulates the growth of hepatocytes and other epithelial cells. A gene for the HGF receptor, c-met, is detected in the intestinal tract and the liver, as well as in gastric carcinoma cells. However, the role of HGF in the regeneration of the normal gastric mucosa is not known. The purpose of the present study was to elucidate the effects of HGF on the morphogenesis of cultured gastric mucosal cells and to evaluate the role of HGF and c-met in the healing process in rat gastric mucosa. The cultured gastric mucosal cells developed a branching morphology in a collagen matrix supplemented with HGF or fetal calf serum. They did not form this morphology on a plastic dish or in the collagen without HGF or the serum. In an in vivo study, total RNA was extracted from rat gastric mucosa 6, 24, 48, and 96 hr after the exposure to a solution of 0.6 M HCl. HGF messenger RNA was not detected, but c-met was expressed in the mucosa. The increased expression of c-met was followed by healing of the mucosal injury. These results indicate that HGF plays important roles in the morphogenesis of gastric mucosal cells and that the HGF receptor gene participates in the healing process of gastric mucosa.

Animals↗
PubMed1995

Colonic mucosal hemodynamics and tissue oxygenation in patients with ulcerative colitis: investigation by organ reflectance spectrophotometry.

Colonic mucosal hemodynamics were investigated at the rectosigmoidal region of the colon in 46 patients with ulcerative colitis and in 18 normal subjects by organ reflectance spectrophotometry under colonoscopy. The value for the index of mucosal hemoglobin concentration (IHb) was significantly higher, and value for the index of mucosal hemoglobin oxygen saturation (ISO2) was significantly lower in patients with active ulcerative colitis than values in the normal controls or in patients with inactive ulcerative colitis. The results indicate mucosal congestion and hypoxemia in patients with active ulcerative colitis. The changes in IHb and ISO2 correlated well with the severity of ulcerative colitis scored by endoscopic findings and with the number of infiltrating inflammatory cells in the mucosa analyzed histologically in biopsy samples. In conclusion, the colonic mucosal microcirculation in patients with active ulcerative colitis was disturbed and showed congestion and hypoxemia. The analysis of hemodynamic changes may be helpful for assessing the activity of ulcerative colitis.

Adult↗