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Biomedical subjects

K Oda

Publications and source records attributed to K Oda.

At least 613 records · Page 34Linked to original sources
PubMed1981

Phasing of nucleosomes in SV40 chromatin reconstituted in vitro.

Phasing of nucleosomes on SV40 DNA was studied by the reconstitution of chromatin from SV40 DNA form I and core histones. The reconstituted chromatin sedimented with increasing S values as the histone : DNA ratios increased, and the buoyant densities in CsCl decreased concomitantly. The average repeat lengths of nucleosomes in the chromatin reconstituted at ratios of 1.0 and 1.5 were estimated to be 168 and 143 base pairs, respectively, by electrophoretic analysis of DNA fragments generated by micrococcal nuclease digestion. The chromatin generated a series of DNA bands that differed in size by about 10 nucleotides upon DNase I digestion followed by heat-denaturation. Phasing of nucleosomes was probed by the use of single-site restriction endonucleases, EcoRI, BamH1, BglI, and HpaII: the latter two cleave DNA at and near the origin of DNA replication and transcription. The form I DNA in the chromatin reconstituted at ratios of 0.5, 1.0, and 1.5 was cleaved up to 60 to 80%, 20 to 60%, and 0 to 10%, respectively. Although the frequency of cleavage by these enzymes was not very different at the ratio 0.5, the BglI site became relatively more susceptible than the other sites at the ratio 1.0. At the ratio 1.5, the DNA was almost resistant to these enzymes, though a significant amount (10%) was cleaved by BglI. These results suggest that the origin is the site unfavorable for nucleosome phasing although the region can be almost completely covered with nucleosomes at higher histone : DNA ratios. The fraction of chromatin immunoprecipitated with anti-T serum after in vitro T antigen binding also decreased with increase in the histone : DNA ratios. The results suggest that T antigen binds preferentially to the internucleosomal region. T antigen preferentially bound to the chromatin reconstituted with the DNA fragment containing the origin. Inefficient phasing of nucleosomes at the origin of DNA replication may facilitate the binding of T antigen to the origin.

Antigens, Neoplasm↗
PubMed1981

Precursor of rat liver alpha 2u-globulin: partial amino acid sequence determination of its signal peptide.

The biosynthesis of alpha 2 u-globulin, the major protein in rat urine, was studied in a cell-free system programmed with messenger RNA isolated from rat liver. Analysis of the primary translation product by SDS-polyacrylamide gel electrophoresis showed a molecular weight of approximatley 22,000 daltons as compared to 20,000 daltons for the mature form. When translation was carried out in the presence of dog pancreas microsomes, alpha 2u-globulin was cotranslationally processed and sequestered in the microsomal lumen. Automated sequencing to mature alpha 2u-globulin and the labeled precursor disclosed the presence of 19 additional amino acids at the amino-terminal end of the primary translation product of the urinary alpha 2u-globulin. The partial amino acid sequence of this extension was determined.

Alpha-Globulins↗
PubMed1981

Multiple forms of rat-serum alpha 1-protease inhibitor. Involvement of sialic acid in the multiplicity of three original forms.

alpha 1-Protease inhibitor was purified from rat serum by two different methods, of which an immunoaffinity method should be preferentially used to obtain all of the multiple forms. The protein thus obtained showed a single protein band in dodecylsulphate/polyacrylamide gel electrophoresis corresponding to a molecular weight of 54000, and contained 13.2% carbohydrate by weight. By column isoelectric focusing in a pH 3.5-5.0 gradient the purified alpha 1-protease inhibitor was separated into five forms with pI values from 4.3-4.7. The amino acid composition of each form was identical, while sialic acid content was significantly different from each other. The most acidic form contained 6.7 residues/molecule, the most basic form, 5.1 residues/molecule, and three forms between them showed proportionally intermediate values between the two. When alpha 1-protease inhibitor was treated with neuraminidase, the five forms were converted finally into three major forms with pI values of 5.3-5.7. In addition, the major form (band 3) of the inhibitor was also converted into three forms after complete removal of sialic acid. These results suggest that alpha 1-protease inhibitor originally exists as three forms with different charges, possibly due to modification of amino acids which might not be detectable by the amino acid composition analysis in the present study. A possible explanation was presented for involvement of sialic acid in appearance of multiple forms originating from three parental forms.

Amino Acids↗
PubMed1981

Myasthenia gravis: passive transfer to mice of antibody to human and mouse acetylcholine receptor.

Mice were given intraperitoneal injections of serum from three patients with myasthenia gravis who had different titers of antibody against mouse acetylcholine receptor (AChR). Almost all mice treated with a high titer of serum antigens showed generalized paralysis, reduced amplitudes of miniature endplate potential (MEPP), or reduced numbers of AChRs. The effects were less marked in serum with lower titers of antibody to mouse AChR, when there was no relation to the severity of effects to the titer of antibody to human AChR.

Animals↗
PubMed1980

Myasthenia gravis: antibodies to acetylcholine receptor with human and rat antigens.

Serum antibodies to acetylcholine receptors (AChR) in myasthenia gravis were surveyed by radioimmunoassay, using 125I-alpha-bungarotoxin-complexed receptors of human muscles and denervated rat muscles. Antibodies were detected more frequently with human AChR than with rat AChR. Furthermore, antibody titers to human AChR were not always parallel to titers observed in the rat receptors, indicating the heterogeneity of antibody specificities in different individuals. Correlation between antibody titers and clinical severity was better when the antibody concentration was determined with human AChR. About 90% of the myasthenic sera contained antibodies to AChR, and 40% also contained other antibodies such as thyroid autoantibodies. These findings suggest that myasthenia gravis is a multiple immunopathy.

Acetylcholine↗
PubMed1979

Purification and properties of a proteinaceous metallo-proteinase inhibitor from Streptomyces nigrescens TK-23.

A novel metallo-proteinase inhibitor which is capable of inhibiting the activities of metallo-proteinases such as the thermolysin, was isolated from the culture filtrates of Streptomyces nigrescens TK-23. The inhibitor was purified batch-wise from the culture filtrate by Amberlite IRC-50 and column chromatographies on CM-Sephadex C-50 and Sephadex G-50. The purified inhibitor showed a single band on 15% polyacrylamide gel electrophoresis at pH 4.3, and at pH 7.5 on SDS-gels. The inhibitor retained 80% of its original activity after treatment of 100 degrees C for 5 min between pH and 7. The molecular weight was estimated to be 12 000 by gel filtration and SDS-polyacrylamide gel electrophoresis, and calcuated as 11 950 from its amino acid composition. The isoelectric point was pH 10.3. The inhibitor showed a high content of hydrophobic amino acids, did not contain tryptophan, and had two disulfide bridges. It also showed specific inhibitory activity for metallo-proteinases but not for serine-, thio- and carboxyl-proteinases.

Amino Acids↗
PubMed1979

Isolation of Golgi fractions from colchicine-treated rat liver. I. Morphological and enzymic characterization.

1. Three Golgi fractions, GF-1, GF-2 and GF-3, were isolated from the livers of rats pretreated with colchicine, which gave better yields of the fractions than ethanol treatment of rats. 2. Electron microscopic observation showed that GF-1 was composed mainly of secretory vesicles, GF-3 consisted predominantly of small tubules and flattened cisternae, and GF-2 was an intermediate fraction composed of secretory vesicles and cisternal elements. 3. Among these three fractions the highest activity of galactosyl transferase, marker enzyme of the Golgi complex, was found in GF-3 and the lowest activity was in GF-1, although a different distribution of the enzymes was observed in fractions obtained from ethanol-treated rat liver. 4. Enzymatic characterization of these fractions showed that no significant contamination with other subcellular components occurred in GF-1 and GF-2.

Acid Phosphatase↗
PubMed1979

Isolation of Golgi fractions from colchicine-treated rat liver. II. Electrophoretic characterization.

1. Intact Golgi fractions, three from colchicine- or ethanol-treated rat livers and two from a control, were analyzed by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. All the fractions showed very similar electrophoretic profiles with 33 protein bands, some of which, especially albumin, had rather higher density in the secretory vesicle fraction than those in the cisternal fraction. 2. Using albumin as the content marker, the Golgi fractions were subfractionated into membranes and contents by freezing-thawing and sonication followed by centrifugation. Distribution of galactosyltransferase among these membrane preparations showed that this enzyme was more enriched in the Golgi cisternal membranes than in the secretory vesicle membranes. 3. All the membrane preparations from the Golgi complex showed very similar patterns on electrophoresis, which were distinctly different from those of microsomal membranes and of plasma membrane. Furthermore, all the Golgi content subfractions had similar protein components, most of which were also found in serum. The microsomal contents, however, showed a considerably different pattern from those of the Golgi contents. 4. From these results it could be concluded that the secretory vesicles are indeed a member of the Golgi complex despite their different appearance and morphology.

Animals↗
PubMed1979

Protease and elastase of Pseudomonas aeruginosa: inactivation of human plasma alpha 1-proteinase inhibitor.

The present study indicates that crystalline elastase of Pseudomonas aeruginosa is a very potent inactivator of human plasma alpha 1-proteinase inhibitor, the enzyme (E) inactivated the inhibitor (I) almost completely within 1 h at 25 degrees C at a molar ratio of E/I = 1:100. The crystalline P. aeruginosa protease also inactivated the inhibitor, but 100-fold less. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the alpha 1-proteinase inhibitor inactivated by the elastase and protease showed decreases in molecular weight of approximately 5,000 and 10,000, respectively. Regeneration of trypsin was negligible even when bovine trypsin-alpha 1-proteinase inhibitor complex (E/I = 1.0) was treated with the elastase. The affinity of alpha 1-proteinase inhibitor to trypsin was much higher than that to elastase. It was suggested that, assuming the pseudomonal proteases are produced and can inactivate alpha 1-proteinase inhibitor in vivo during pseudomonal diseases, the loss of alpha 1-proteinase inhibitor activity may permit the endogenous serine proteases to cause tissue destruction.

Humans↗
PubMed1979

Pepstatin-insenstive acid proteases from Scytalidium lignicolum. Kinetic study with synthetic peptides.

A kinetic study was conducted on the acid proteases A-1 and A-2 from Scytalidium lignicolum using synthetic peptides as substrates. Almost maximum activity was attained with N-acylated tetrapeptides as the molecular size of substrates was increased. Suitable amino acid residues were required at the P1-P2 and P1'-P2' positions [notation of Schechter and Berger (14)]. Hydrophobic or bulky residues such as leucine were specifically required at the P1 and P1' positions, with the specificity at the latter position being considerably lower than that at the former. For catalysis, the presence of certain amino acid residues at the P2 and P2' positions was essential, mainly in relation to kcat. An inhibition study supported this view. Stringent stereospecificity was observed at the P2 and P2' positions, but the side chain specificity was low. Study of the B enzyme from the same organism was very difficult owing to its low activity against the peptides used. The Scytalidium acid proteases A-1, A-2, and B showed considerably different behavior against peptide substrates in comparison with usual acid proteases, which are senstive to pepstatin.

Aspergillus niger↗
PubMed1979

Hematological and biochemical values of thoroughbred foals in the first six months of life.

Hematological and biochemical parameters in five Thoroughbred foals during the first six months of life are reported. The samples were analyzed for red blood cell, packed cell volume, hemoglobin, platelet, white blood cell, absolute number for leukocytes, and erythrocyte fragility and serum calcium, inorganic phosphorus, sodium, potassium, chloride, magnesium, alkaline phosphatase, icterus index unit, bilirubin, blood urea nitrogen, nonprotein nitrogen, blood glucose, lacticdehydrogenase, glutamic pyruvic transaminase, glutamic oxalacetic transaminase, total protein, albumin, globulin, and A/G. The changes of these parameters are presented in relation to age after birth.

Alanine Transaminase↗
PubMed1978

Protein kinases and their protein substrates associated with chromatin and ribosomes in SV40-transformed rat cells.

The level of endogenous protein phosphorylation in non-histone chromosomal and ribosomal wash proteins is 7--10 times greater in SV40-transformed rat cells than in untransformed parental cells. Protein kinase activity in these proteins was fractionated by either phosphocellulose or DEAE-cellulose chromatography. One major and one minor component were detected in non-histone proteins and only one component in ribosomal wash proteins when the activity in each fraction was measured with an exogenous substrate, casein. These enzymes prefer casein to whole histone as substrate and are cyclic AMP-independent. The enzyme activity in a major peak of non-histone proteins and in ribosomal wash proteins measured with casein as substrate is 3 times greater in transformed cells than in untransformed cells, whereas pH optimum, cation requirements and apparent Km values for casein and ATP are identical or very similar in the two cell types. No significant phosphatase was detected in non-histone and ribosomal wash proteins from the two types of cell. The patterns of endogenous protein phosphorylation in these protein fractions analysed by gel electrophoresis are significantly different between these cells. These results suggest that the high level of endogenous protein phosphorylation in non-histone and ribosomal wash proteins from SV40-transformed cells is caused mainly by the increased activity of protein kinase and the nature of protein substrates.

Animals↗