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Y Matuo

Publications and source records attributed to Y Matuo.

At least 55 records · Page 3Linked to original sources
PubMed1984

Biochemical difference between ventral and dorsolateral prostates: effects of castration and administration of the antiandrogen 16 beta-ethyl-17 beta-hydroxy-4-oestren-3-one on SDS-electrophoretic patterns of subcellular proteins from rat prostates.

Protein species in the cytosol, postnuclear particulate, and nuclear fractions from the ventral and dorsolateral prostates of rats castrated or injected with antiandrogen, 16 beta-ethyl-17 beta-hydroxy-4- oestren -3-one ( EHOO ), were analyzed by SDS-polyacrylamide gel electrophoresis. The effects of EHOO injection on the contents of the most abundant species in all three fractions of ventral prostate were similar to the effects of castration, but the effects of EHOO and castration were only similar on the nuclear fraction in the dorsolateral prostate. The most sensitive species to castration in the three subcellular fractions of dorsolateral prostate were the same as those to EHOO injections; this was not the case for the cytosol and nuclear fractions in the ventral prostate.

Androgen Antagonists↗
PubMed1982

Affinities of various nucleases to DNA-Sepharose under non-digestive conditions: survey for productive affinity chromatography.

1. It has been reported that DNase I can be highly purified from pancreas extract by affinity chromatography on a dDNA-Sepharose column under non-digestive conditions. In the present study, the adsorption-elution of other nucleases on the column under non-digestive conditions was studied. 2. All the seven kinds of nucleases tested were adsorbed when applied on a dDNA-Sepharose column under conditions which did not allow the enzymes to hydrolyze the DNA. The non-digestive conditions were as follows. i) For DNase II (pI=10.2), pH 3.0 in the presence of 50 mM sodium sulfate (inhibitor), ii) for micrococcal nuclease (pI=9.6), pH 4.0 in the absence of Ca2+ (activator), iii) for restriction endonucleases Eco RI (pI=5+1), Hind III (pI=5+1), and Bam HI (pI=5+1), pH 4.0 in the presence of 20% glycerol and 0.1% Neopeptone (stabilizers), and iv) for nucleases S1 (pI=5+1) and nuclease P1 (pI=4.5), pH 7.0. At the respective pH's, the enzymes other than nucleases S1 and P1 were cationic so as to exhibit electrostatic attraction to the anionic dDNA-Sepharose. Although S1 and P1 were anionic, they still adsorbed to the column. 3. All the adsorbed nucleases described above were eluted by a concentration gradient of KCl without changing pH. The ionic strengths required for elution were 0.19 for DNase II, 0.53 for micrococcal nuclease, 0.73 for Eco RI, 0.72 for Hind III, 0.37 for Bam HI, 0.17 for P1, and 0.13 for S1. The fact that the ionic strength required for the elution of DNase I (pI=5.0) was 0.39 at pH 4.0 indicates that the former five enzymes except DNase II can be chromatographed with almost the same or higher efficiency than DNase I, because the proteins adsorbed with no-specific affinity could be mostly eluted at lower ionic strength. On the other hand, the fact that nucleases P1 and S1 were adsorbed in spite of electrostatic repulsion suggests that these two enzymes can also be effectively chromatographed, especially when other cationic proteins are previously removed by an appropriate method such as adsorption to a typical cation exchanger.

Animals↗
PubMed1982

Effect of partial hepatectomy and tumor-bearing on phosphorylation of nuclear protein in rats.

When nuclei from various tissues of rats were incubated at 25 degrees with ATP[gamma-32P], the amounts of 32P incorporated in histone and non-histone protein per mg DNA reached maxima at 1 hr (maximum phosphorylation level). H1 histone and over 30 species of non-histone protein were phosphorylated as detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Only some of those species of non-histone protein, which were so low in content as to be indetectable by conventional protein staining, were prominently phosphorylated; their apparent molecular weights were 106K, 82K, 71K, 62K, 39K, 38K and 28K with brain, 39K, 38K, 32K, 31K and 22K with kidney and liver, 17.5K and 11.5K with thymus, and 11.5K with Rhodamine sarcoma (tissue-specific). In phosphorylation with liver nuclei, the tissue specificity changed neither with partial hepatectomy nor upon transplanting Rhodamine sarcoma. However, the maximum phosphorylation levels of the five species of non-histone protein increased with operation and decreased with tumor-bearing, to similar extents. On the other hand, the amounts of all the species of histone per mg DNA changed neither with operation nor with tumor-bearing, whereas the maximum phosphorylation level of H1 histone increased with operation, but not with tumor-bearing.

Animals↗
PubMed1981

Purification and characterization of alkaline protease and neutral protease from chromatin of rats.

It was previously reported that, in addition to a known chymotrypsin-like protease capable of hydrolyzing histones with an optimum pH of 8 (neutral protease), another protease is bound to the chromatin of various rat tissues and in situ hydrolyzes casein more quickly than histones with an optimum pH of 10 (alkaline protease). In the present study, the alkaline protease was purified 14 000-fold to approx. 75% purity from the chromatin of Rhodamine sarcoma. This tumor contains both proteases at higher levels than normal tissues. For purification, affinity columns of Sepharose with bound soybean trypsin inhibitor, casein and histones were successively used. Also, the neutral protease was purified 920-fold to an apparently homogeneous state by affinity chromatography on a Sepharose column with bound soybean trypsin inhibitor under conditions, in which an excess amount of the enzyme was applied on the column so that part of the enzyme would pass through the column without adsorption and the enzyme thus adsorbed was then eluted. The purified alkaline and neutral proteases had molecular weights of approx. 18 000 and 27 000, respectively, and isoelectric points of approx. 11. The former enzyme hydrolyzed casein (100) in preference to histones (18) with an optimum pH of 9.5, whereas the latter enzyme preferred histones (100) to casein (32) with an optimum pH of 8. Their actions against other proteins and synthetic substrates were also studied.

Animals↗
PubMed1981

Tumor-associated serum proteins in rats as detected by two-dimensional gel electrophoresis.

The sera from normal and tumor-bearing rats were separated by polyacrylamide gel disc isoelectric electrophoresis and then in a perpendicular dimension by polyacrylamide gel slab electrophoresis with a gradient from 4% to 20%. When the gel was stained with Coomassie brilliant blue, 100-150 components were separately observable with normal serum. With the sera from Rhodamine sarcoma-bearing rats, three more components were detected with a high reproducibility. They were of pI (isoelectric point) 4.5 and Rm (relative mobility) 0.63, of pI4.8 and Rm 0.63 and of pI 4.9 and Rm 0.75. The 4.5/0.63 (pI/Rm) and the 4.8/0.63 components appeared soon after transplanting the tumor, whereas the 4.9/0.75 component appeared at the late stage of its growth. Upon surgical removal of the grown tumor, the former two components disappeared rapidly, while the latter component disappeared gradually. Similar results were obtained with Yoshida sarcoma, and ascites hepatoma AH66 and AH130. A 3rd polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate revealed that the 4.5/0.63 and the 4.8/0.63 components were each composed of a single polypeptide of approximately 64,000 daltons, and that the 4.9/0.75 component was composed of two different polypeptides of approximately 33,000 daltons and approximately 10,000 daltons. It was confirmed by an immunodiffusion experiment that none of these three proteins was alpha-fetoprotein.

Animals↗
PubMed1980

Affinity chromatography of porcine pancreas deoxyribonuclease I on DNA-binding sepharose under non-digestive conditions, using its substrate-binding site.

1. DNase I from porcine pancreas, if Mg2+ was present, hydrolyzed both sDNA and dDNA, whether free or bound to Sepharose. The hydrolysis rates were maximum at pH 7.5 with the bound DNAs and at pH 7.0 with the free DNAs negligible at pH 4.0 and pH 10.5 with the free and bound DNAs. The hydrolysis was completely inhibited by 50 mM sodium citrate. 2. With 50 mM citrate buffer (Ph 4.0), DNase I was effectively adsorbed on the DNA-Sepharoses in the absence of 5 mM Mg2+. The adsorbed enzyme was effectively eluated by the buffer containing 1 M KCl (eluate). The amounts of the eluated enzyme were approximately 1.5 X 10(5) units/mg DNA with sDNA-Sepharose and approximately 3.0 X 10(5) units/mg DNA with dDNA-Sepharose. This simple adsorption-elution of the pancreas extract resulted in approximately 300-fold purification of DNase I with a yield of 95%. In the elute, the ratios in activity of trypsin, chymotrypsin and RNase to DNase I were 1/(4.0 X 10(5)), 1/(5.3 X 10(3)), and 1/(4.1 X 10(2)) as low as in the extract, respectively. In addition, the eluate was not contaminated by kallikrein or carboxypeptidases A and B. 3. Upon repeating the adsorption-elution described above, the adsorbing capacities of DNA-Sepharoses gradually deteriorated with the whole pancreas extract, but not with the precipitate of the extract formed on 60% ammonium sulfate saturation, which contained 90% of the DNase I. With the precipitate, one dDNA-Sepharose solumn was repeatedly usable at least 20-times without deterioration. The DNase I preparation thus obtained was homogeneous on SDS-polyacrylamide gel electrophoresis. 4. Conceivably, the above-mentioned adsorption of DNase I on DNA-Sepharoses was mainly due to the steric and electrostatic affinity of a relatively large moiety of the DNA molecule to the substrate-binding site, but not to the catalytic site, of the enzyme.

Animals↗
PubMed1976

Small fragments from the A subunit of cholera toxin capable of activating adenylate cyclase.

Exposure of cholera toxin to membrane particles prepared from sarcoma 180 cells gives rise to a variety of fragments which are capable of activating adenylate cyclase [ATP:pyrophosphate-lyase (cyclizing), EC 4.6.1.1]. A major component of these fragments has an apparent molecular weight in the 8,000-10,000 range. The smallest stimulatory fragment has a molecular weight of approximately 1400. The small size of the fragments is confirmed by Sephadex gel filtration, in the presence of either sodium dodecyl sulfate or formic acid. These fragments are produced from holotoxin or its A subunit by protease(s) found in sarcoma membrane particles. Production of fragments appears optimal in 40-60 min at 30 degrees and pH 7, and is prevented by protease inhibitors. The ability of the small fragments to activate adenylate cyclase is reversed by anti-holotoxin, but not anticholeragenoid, antibodies. These fragments require NAD for the activation of adenylate cyclase and are fully active after heating at 90 degrees for 5 min (pH 7).

Adenylyl Cyclases↗