巻号一覧
62 巻, 6 号
選択された号の論文の17件中1～17を表示しています
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I. Nature and Properties of the Proteolytic Enzyme SystemYOSHIKO NARAHASHI, MASAYA YANAGITA1967 年 62 巻 6 号 p. 633-641
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーThe occurrence and properties of several kinds of endopeptidase and exopeptidase in the commercially available proteinase preparation of Streptomyces griseus K-1, Pronase, were investigated.
1. It was found that Pronase contains at least two kinds of peptidase in addition to proteinases.
2. A study of the action of the two peptidases indicated that one of them hydrolyzed LG and LNA like an aminopeptidase and the other hydrolyzed CGL like a carboxypeptidase, and both activities remarkably increased in the presence of Ca and Co ions.
3. The two peptidases were both metalloenzymes, and the binding of activating metal with EDTA rendered the enzymes inactive. Upon reactivation by Co and Ca ions, both activities were restored.
4. The peptidase hydrolyzing LG and LNA was found to be rather heat stable below 80°C. Moreover, it was not inactivated by up to 9M urea even at room temperature.In contrast, it was very labile on dialysis against distilled water and with metal-chelating reagents. With LG the pH optima were around pH 8.3 in the presence of Ca ion and at pH 8.0 and 9.5 in the presence of Co ion.
5. The peptidase which hydrolyzes CGL was less heat stable. However, it was stable on dialysis against distilled water. The pH optimum was at pH 7.5 in the presence of Co or Ca ion.
6. About 70 per cent of the caseinolytic activity derived from proteinases in Pronase were irreversibly inhibited by EDTA treatment. The remaining 30 per cent of the caseinolytic activity contained all the esterase activity for BAEE. With casein, the proteinases were optimally active over a wide pH range of 7.0 to 9.0 like the esteriolytic activity for BAEE.抄録全体を表示PDF形式でダウンロード (1534K) -
XII. Biochemical Studies (7). Dephospho-CoA Pyrophosphorylase and Dephospho-CoA Kinase as a Possible Bifunctional Enzyme ComplexTADAO SUZUKI, YASUSHI ABIKO, MASAO SHIMIZU1967 年 62 巻 6 号 p. 642-649
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリー1. Dephospho-CoA pyrophosphorylase [EC 2.7.7.3] and dephospho-CoA kinase [EC 2.7.1.24] were purified in the same fraction 248-and 266-fold, respectively, from rat liver 59, 000×g supernatant. The ratio of specific activities of both enzymes remained constant throughout the purification procedures.
2. These two enzymes were not separated each other by CMcellulose chromatography, DEAE-cellulose chromatography and Sephadex G-200 gel filtration.
3. Inactivation by heat and stimulation by magnesium ion proceeded in parallel with these two enzyme activities.
4. In the presence of 0.01%of deoxycholate, the dephospho-CoA pyrophosphorylase reaction was stimulated, whereas dephospho-CoA kinase reaction was inhibited. Preincubation with 0.2%deoxycholate caused different changes in each of the two enzyme activities.
5. From the results obtained, a possible bifunctional enzyme complex system of these two enzymes was discussed.
6. The Michaelis constants of dephospho-CoA pyrophosphorylase for D-pantetheine-4'-phosphate and for ATP were 1.4×10-4M and 1.0×10-3M, respectively, and those of dephospho-CoA kinase for dephospho-CoA and for ATP were 1.2×10-4M and 3.6×10-4M, respectively.抄録全体を表示PDF形式でダウンロード (1401K) -
CI. 12α-Hydroxylation of Chenodeoxycholic Acid in Eel LiverTAKAKO MASUI, FUJIKO UEYAMA, HIROMASA YASHIMA, TARO KAZUNO1967 年 62 巻 6 号 p. 650-654
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーThe in vivo and in vitro conversion of chenodeoxycholic acid in the eel was studied.
1. Chenodeoxycholic acid-3H injected intraperitoneally into the eel was efficiently converted into cholic acid-3H.
2. Chenodeoxycholic acid-3H was hydroxylated to cholic acid-3H by 12α-hydroxylase activity in the microsomal fraction of eel liver in vitro.抄録全体を表示PDF形式でダウンロード (791K) -
NOBUHIKO HOSHITA, KYUICHIRO OKUDA1967 年 62 巻 6 号 p. 655-657
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーPartial syntheses of 3α, 7α-dihydroxy-5β-cholan-24-al through Grundmann aldehyde synthesis and of 3α, 7α-dihydroxy-5β-cholest-24-en-26-oic acid through Wittig reaction were described.抄録全体を表示PDF形式でダウンロード (549K) -
MICHIHIKO TADA1967 年 62 巻 6 号 p. 658-672
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーThe changes in the filament structures of dog cardiac myosin B induced by ATP were studied by electron microscopy. At higher ionic strengths a polarized “arrowhead” structure was seen, which disappeared on addition of ATP, while the ordinary actin structure appeared. At lower ionic strengths, myosin aggregates were seen besides actin filaments in the presence of a high concentration of ATP, where myosin B showed a clearing response, while in the presence of a low concentration of ATP, where myosin B showed superprecipitation, many large aggregates were formed.
The kinetics of the ATPase and superprecipitation of dog cardiac myosin B were investigated under different conditions. The extent and rate of superprecipitation of cardiac myosin B induced by ATP remained at constant levels above a critical myosin B concentration. Below this the extent was proportional to the concentration of myosin B, but the specific rate decreased with decrease in the myosin B concentration and superprecipitation did not occur below a critical protein concentration. Both the rate of superprecipitation and the ATPase activity of cardiac myosin B decreased with increasing KC1 concentration up to a critical concentration, above which superprecipitation did not occur and the ATPase activity decreased to a level almost equal to that of myosin alone. The minimum concentration of ATP required to maintain the maximum extent of superprecipitation decreased on addition of pyruvate kinase and phosphoenolpyruvate, and the minimum amount of ATP required in the presence of a sufficient amount of the kinase was about one mole, or less than one mole, per mole of cardiac myosin. Over a wide range of ATP concentration, the ATPase activity and superprecipitation of cardiac myosin B were sensitive to Ca++ and EGTA, and the ATPase activity and the extent and rate of superprecipitation increased as the ATP concentration increased until a critical concentration was reached. Above this the clearing response and the substrate inhibition of ATPase took place. The substrate inhibition of cardiac myosin B ATPase was less marked than that of the skeletal myosin B ATPase, particularly in the presence of Ca++, and the clearing response was often accompanied by a high actomyosin type ATPase activity. Even in the presence of a large excess of ATP, where myosin B showeda typical clearing response, the ATPase activity of cardiac myosin B was several times greater than that of cardiac myosin alone in the presence of either Ca++ or EGTA. With aging of the preparation of cardiac myosin B, the superprecipitation and ATPase activity were very rapidly desensitized to Ca++ and EGTA, and the critical concentration of ATP required to induce the substrate inhibition of the ATPase of aged cardic myosin B was markedly higher than that for a fresh preparation. In the presence of the minor component of metin from skeletal muscle, the ATPase activity and superprecipitation of an aged preparation of cardiac myosin B were sensitized to Ca++ and EGTA. The kinetics of the ATPase of cardiac myosin, viz. the constituent of myosin B, was also studied. The amount of the initial extra-liberation of Pi from the cardiac myosin-ATP system was about one mole per mole of myosin. At the steady state the cardiac myosin ATPase reaction followed the Michaelis-Menten kinetics, and the Michaelis constant and the maximum rate for cardiac myosin ATPase in 0.5 M KC1-2mM MgCl2-0.8mM phosphoenolpyruvate-0.15mg. pyruvate kinase per ml. at pH 7.0 and 25°C were found to be 3.5μM and 2.6μmoles g.-1 min. -1, respectively.抄録全体を表示PDF形式でダウンロード (4669K) -
II. Effect of Terminal Phosphate of Ribonucleic Acid and Effect of Ribonucleoside 5'-DiphosphateKENJI ISHII, SATORU SHIMIZU, ISAMU SHIIO1967 年 62 巻 6 号 p. 673-678
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーCauses for the low reactivity of yeast RNA as compared with Poly A in the degradation by polynucleotide phosphorylase [EC 2.7.7.8] from Azotobacter agilis were studied. Since the modified commercial RNA treated with phosphatase [EC 3.1.3.1] was phosphorolyzed and arsenolyzed eighteen and sixteen times, respectively, faster than the untreated commercial RNA and since the initial rate obtained for the treated. RNA was even higher than that obtained for Poly A, the phosphomonoester end groups of the RNA were considered to be mainly responsible for the low reactivity. The rapid decrease of reaction rate in the later stage of the degradation of commercial RNA was ascribed to the product inhibition. The product, ADP, GDP or CDP (each 0.3mg./ml.) and a mixture of ADP, GDP, CDP and UDP (each 0.3mg./ml.) caused 15% and 60% inhibition, respectively, of the phosphorolysis of dephosphorylated commercial RNA (1.0mg./ml.). The RNA with a terminal phosphate, which remained unattacked, also inhibited the degradation. On the other hand, the degradation rate of highly polymerized RNA was not affected by the phosphatase treatment. The hydrogen bond in the structure of this RNA seems, therefore, to be responsible for the low degradation rate.抄録全体を表示PDF形式でダウンロード (1112K) -
TOMOYUKI SHINOMIYA, FUJIO EGAMI1967 年 62 巻 6 号 p. 679-687
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリー1. Preferential inhibition of pyocin R production by dihydrostreptomycin (DHSM) was confirmed and further investigated.
2. Synthesis of pyocin R protein was remarkably inhibited by DHSM at 5μg./ml., whereas little decrease of total protein synthesis was observed.
3. The existence of a stage especially sensitive to DHSM in the process of pyocin R production was suggested.抄録全体を表示PDF形式でダウンロード (1710K) -
III. Fate of Parental Phage DNA and Replication of the RFKENICHI MATSUBARA, KAZUNORI SHIMADA, YASUYUKI TAKAGI1967 年 62 巻 6 号 p. 688-699
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーDensity studies have been carried out to follow the process of RF (replicative form) production and multiplication in E.coil 15T-infected with bacteriophage ∅X174.
It has been shown that the injected single-stranded phage DNA is either converted to double-stranded parental RF or brokendown and reutilized for host DNA synthesis.The conversion process to make parental RF is carried out by de novo synthesis of the complementary strand.It is not carried out by interaction of the viral DNA with preformed bacterial materials of high molecular weight.
Upon subsequent incubation of the infected complex, replication of RF occurs and many progeny RF both strands of which are synthesized de novo after infection are accumulated. The mode of replication of the RF is semiconservative. Evidence is presented which strongly suggests that during the replication process, not only parental RF but also progeny RFs undergo replication抄録全体を表示PDF形式でダウンロード (2607K) -
Purification, properties and Application in Carbohydrate ResearchTAKASHI MURAMATSU, FUJIO EGAMI1967 年 62 巻 6 号 p. 700-709
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーThe liver of Turbo cortunus contained several glycosidase activities. Sephadex G-200 column chromatography, heating and hydroxylapatite column chromatography were effective for the separation of the glycosidases from each other.a-Mannosidase [EC 3.2.1.24] from the liver of T.cortunus was purified 130-fold, using p-nitrophenyl α-D-mannoside as substrate.The purified a-mannosidase was practically free from all other glycosidases tested. The enzyme released 64-66% of the mannose from ovalbumin glycopeptide.It also released mannose from pronase digest of Taka-amylase A [EC 3.2.1.1] and a mannosyl rhamnose, which is a part of the repeating oligosaccharide unit of the o-antigenic lipopolysaccharide in Salmonella typhimurium.The enzyme did not attack yeast mannan.
β-Mannosidase [EC3.21.25] from the livcr of T.cortunus was purified 63-fold, using phenyl β-D-mannoside as substrate. The purified β-mannosidase was practically free from all other glycosidases tested. The enzyme hydrolyzed none of the four compounds mentioned above.抄録全体を表示PDF形式でダウンロード (1528K) -
I. Purification and Enzymatic Activity of Cytochrome Oxidase from Saccharomyces oviformis M2ICHIRO SEKUZU, HIROSHI MIZUSHIMA, SHUNTARO HIROTA, TOSHITSUGU YUBISUI, ...1967 年 62 巻 6 号 p. 710-718
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーCytochrome oxidase [EC 1.9.3.1] was isolated and purified from ETP of Sacchromyces oviformis M2 by means of cholate and ammonium sulfate extraction and ammonium sulfate fractionation. The purified preparation contained approximately 7-8mμ moles heme α and copper per mg. protein.
The activity of the purified preparation was about 20 fold more than that of ETP. The optimum pH was found at 6.2 and the optimum phosphate buffer concentration was 67mM. The Km value of cytochrome c was 2-4×10-6M. The maximum turnover number of the purified preparation was about 14, 000 which was 10 fold higher than that of a beef heart preparation.
Based on the above results together with other properties, the properties of cytochrome oxidase of baker's yeast were discussed.抄録全体を表示PDF形式でダウンロード (1411K) -
MASANOBU TOKUSHIGE, IRWIN W. SIZER1967 年 62 巻 6 号 p. 719-725
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーGlycolate oxidase [EC 1.1.3.1, glycolate: oxygen oxidoreductase, kidney] was purified 740 fold from hog kidney renal cortex by calcium phosphate gel treatment, ammonium sulfate fractionation and column chromatography on DEAE-cellulose and hydroxylapatite.The glycolate oxidase utilized glycolic acid most rapidly, and DL-glyceric, α-hydroxybutyric and L-and D-lactic acids much more slowly. The Km, for glycolate was 0.30mM. FMN was found to be a cofactor of the enzyme, as judged by the absorption spectrum, activating effect, and paper electrophoretic analysis. The Km, for FMN was 0.71mM
Employing a newly devised microassay method including solvent extraction of radioactive compounds, glyoxylic acid was shown to be the enzymatic product of the oxidase reaction, but not oxalic acid; in the absence of catalase [EC 1.11.1.6] glyoxylic acid was further oxidized nonenzymatically to yield formic acid and CO2 by H2O2, which was generated during the oxidase reaction.抄録全体を表示PDF形式でダウンロード (1365K) -
I. Properties of the FactorYASUYUKI KOZUKI, YUJI TONOMURA1967 年 62 巻 6 号 p. 726-739
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーA regulatory factor, which changes dramatically and reversibly the state of aggregation of myosin, was found in the metin preparation of Szent-Györgyi and Kaminer.The factor, named “myosin aggregation factor” (MAF), was inactivated at 60°C, and precipitated at pH 5.0. It was soluble in 0.1M KCI at pH 7.0, was salted out from 20 to 55 per cent saturated ammonium sulfate, and was precipitated in 0.1M KCI by ultracentrifugation at 35, 000 r.p.m.for 3 hours.When the solution from the precipitate was dialyzed against an ATP solution, it showed the myosin aggregation activity only slightly, even after the addition of 0.1M KCI and 2 mM MgCl2.Its ultraviolet absorption spectrum exhibited a peak at 280mμ.The factor lost its activity by digestion with Pronase-P, but not with RNase-A[EC 2.7.7.16], indicating that the factor is a protein.
Myosin and MAF formed a complex, which in the absence of ATP showed spherical aggregates in 0.1M KCl and 2 mM MgCl2 at pH 7.0. These aggregates were transformed into a net-work of filaments upon addition of ATP.After splitting the ATP, the complex formed again the spherical aggregates.Magnesium ion and a minute amount of calcium ion had to be present in order to induce the above transition.
By measuring the effect of the supernatant obtained by centrifuging a mixture of myosin and MAF preparation on the aggregation state of myosin and its protein content, it was concluded that the amount of MAF binding with myosin is about 10 per cent by weight of myosin, while its aggregation effect is already saturated by the addition of only about 1 per cent MAF by weight of myosin.
H-Meromyosin in 0.1M KCI was insolubilized by the addition of MAF. The effect was saturated by the addition of 4 per cent MAF by weight of H-meromyosin.The complex of H-meromyosin and MAF was solubilized on the addition of ATP, and became insoluble again after ATP was split.抄録全体を表示PDF形式でダウンロード (10341K) -
SHOGO ICHII, NAGASUMI YAGO, SHIGERU KOBAYASHI, SABURO OMATA1967 年 62 巻 6 号 p. 740-745
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリー1. Fresh mitochondria from hog adrenal cortex contained a high concentration of P-450 but no detectable P-420.
2. Thirty-fold purification of cholesterol 20α-hydroxylase from acetone-dried mitochondria was achieved, but no definite tendency of increase in P-450 concentration was observed during the enzyme purification.
3. 20α-Hydroxylase activity and P-450 in purified preparations decreased significantly during aging at 0°C, whereas P-420 retained the original level. Glycerol, ethylene glycol and propylene glycol prevented the degradation of P-450 but not of P-420 and protected 20α-hydroxylase from inactivation by aging.
4. At low concentrations (up to 10μM) FAD enhanced 20α-hydroxylase activity but at higher concentrations it inhibited the enzyme activity considerably.
5. The enzymatic properties of the cholesterol 20α-hydroxylase complex were discussed.抄録全体を表示PDF形式でダウンロード (1129K) -
SHIGERU MATSUZAKI, MITSUO SUZUKI1967 年 62 巻 6 号 p. 746-755
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーStudies were carried out on the metabolism of iodinated amino acids in the pig thyroid with the following results:
1. It was confirmed that NADPH-dependent, iodotyrosine-deiodinating activity is localized mainly in the microsomal fractions.
2. L-Monoiodotyrosine was revealed to be present as an intermediate during the deiodination of L-diiodotyrosine. This fact, together with oxygen independency and requirement of reduced nicotinamide nucleotide for this reaction, indicates that the deiodination of iodotyrosines is of a reductive nature.
3. Iodotyrosine deiodinase is highly specific for L-iodotyrosines. At the substrate concentration of 1×10-6M, the rate of deiodination of Ldiiodotyrosine was about one fifth the rate of deiodination of L-monoiodotyrosine. At the optimal pH of 8.0, the Km for L-diiodotyrosine was greater than that for L-monoiodotyrosine, the maximal velocity being the same for each iodotyrosine.
4. Some other enzymatic properties of the enzyme such as cofactor requirement, and sensitivity toward inhibitors were investigated.
5. L-Tyrosine, and keto acid analogues of monoiodotyrosine and diiodotyrosine were demonstrated to act as competitive inhibitors of the deiodination reaction.
6. Peroxidase [EC 1.11.1.7], NADPH-cytochrome c reductase and DT-diaphorase [EC 1.6.99.2], each derived from pig thyroid, do not appear to be concerned in the deiodination of iodotyrosines.抄録全体を表示PDF形式でダウンロード (1782K) -
AKIO YOSHIMOTO, TARO NAKAMURA, RYO SATO1967 年 62 巻 6 号 p. 756-766
発行日: 1967/12/25
公開日: 2011/06/07
ジャーナル フリーA protein catalyzing the reduction of sulfite and hydroxylamine by reduced viologen dyes was purified from Aspergillus nidulans to an ultracentrifugally homogeneous state. No physiological electron donors. including NADH and NADPH. could be utilized by this protein for sulfite reduction. The purified protein had a sedimentation coefficient of 4.23S and showed a spectrum having peaks at 384 and 585mμ and a shoulder at 453mμ. When hydrogen and Desulfovibrio hydrogenase were used to generate reduced methyl viologen (MVH). this protein reduced sulfite quantitatively to sulfide according to the equation: SO3--+3H2→S--+3H2O. The sulfite-reducing activity was inhibited by cyanide, arsenite.and sulfide.A mutant strain of A. nidulans incapable of assimilating sulfate and sulfite did not contain the MVH-sulfite reductase activity.抄録全体を表示PDF形式でダウンロード (3251K) -
HIROKO TODA, KOZO NARITA1967 年 62 巻 6 号 p. 767-768
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーPDF形式でダウンロード (321K) -
YOSHIHO NAGATA, BUNJI MARUO1967 年 62 巻 6 号 p. 769-771
発行日: 1967/12/25
公開日: 2011/01/25
ジャーナル フリーPDF形式でダウンロード (638K)
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