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60 巻, 1 号
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HIROSHI TERAYAMA, AYAKO TAKATA1966 年 60 巻 1 号 p. 1-11
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリー1. Whole body irradiation of rats with Co60-γ rays did not affect either the aminoazo dye N-demethylating activity or the hydro-xylating activity of rat-liver. It also had no significant effect on the induction of the enzymatic activities by methylcholanthrene. The ionizing irradiation, however, appeared to increase the reduction of enzymatic activity following partial hepatectomy.
2. The time course of the N-demethylat-ing activity of regenerating rat liver after partial hepatectomy or a sham operation (laparotomy) was studied. In both cases, a remarkable reduction (50%) in the enzymatic activity was observed within a few hours after the operation. In the case of sham operation, the activity returned to the normal level within one day. In the case of partial hepatectomy, however, the activity remained at a low level for 24 hours and then later gradually returned to the normal level over a period of four days and in parallel with liver regeneration.
3. Administration of cortisone had no significant effect on the N-demethylating activity of normal liver but partially prevent-ed the decrease in enzymatic activity caused by adrenalin injections or partial hepa-tectomy.
4. Administration of adrenalin caused a large and a rapid decrease in N-demethylat-ing activity. This was also observed in methylcholanthrene-treated rats.
5. Methylcholanthrene was shown to induce N-demethylating activity in partially hepatectomized rats. A large prior dose of methylcholanthrene appeared to prevent reduction in the enzymatic activity after partial hepatectomy.
The authors wish to express their sincere thanks to Mr. Mitsuo Matsumoto and Dr. Tokutaro Hishizawa for their co-operation and valuable discus-sions throughout the study.抄録全体を表示PDF形式でダウンロード (718K) -
KAZUO SATAKE, TERUO TAKE, ATSUKO MATSUO, KOSHI TAZAKI, YOICHI HIRAGA1966 年 60 巻 1 号 p. 12-16
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリーAn analyzer for the quantitative deter-mination of amino acids and peptides with 2, 4, 6-trinitrobenzenesulfonic acid is described. The instrument could be used both for rapid quantitative monitoring of chromatographic effluents and for discontinuous analysis of samples collected previously.
The sensitivity was rather high and 0.03 to 0.3 μmoles of amino acids could be assayed with an accuracy of ±4 per cent. After the colorimetry TNP-amino acids and -peptides could be recovered and readily reconverted to the original compounds.抄録全体を表示PDF形式でダウンロード (365K) -
V. The Effects of Metal Ions and Sulfhydryl Reagents on the Absorption Spectrum of the EnzymeEIICHI MISAKA, KAZUO NAKANISHI1966 年 60 巻 1 号 p. 17-26
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリーThe changes in the absorption spectrum of lipoamide dehydrogenase from Saccharomyces oviformis were investigated on addition of excess NADH2 in the presence of various metal ions, arsenite or PCMB under anaerobic conditions.
With group I metal ions (Fe++ and Co++), the flavin moiety of the enzyme is reduced by NADH2 to a red-intermediate and is oxidized in air to a yellow form. This suggests that the metal combines weakly with the dithiol group of the enzyme and is split from the enzyme protein by reoxidation in air. With group II metal ions (Zn++, Cd++ and Cu++), the enzyme is reduced by NADH2 to a green form via a red-intermediate and shows a new absorption band at a longer wavelength. On reoxidation in air, the green form changes to a yellow form, the spectrum (band around 450 mμ) of which is shifted to a shorter wavelength. After BAT.L treatment, the spectrum returns to that of native enzyme. Arsenite also combines with the dithiol group of the enzyme which has been fully reduced by NADH2 via a red-intermediate. The green form shows a new absorption band at a longer wavelength and is reoxidized in air to the yellow oxidized enzyme, suggesting the dissociation of arsenite from the enzyme protein. Like native enzyme, PCMB-treated enzyme in which free -SH groups are blocked, is reduced by NADH2 to a red-intermediated. This PCMB-enzyme is reduced by NADH2 in the presence of PCMB to a colorless form via a red-intermediate. It has no absorption band at a longer wavelength. It changes to a yellow oxidized form in air, with a spectrum shifted to shorter wavelengths. This reoxi-dized enzyme is directly reduced by NADH2 to the colorless form. In the presence of Cd++, the PCMB -enzyme is reduced by NADH2 to a green form via a red-inter-mediate. The spectrum of the green form shows no absorption band at a longer wave-length. On reoxidation in air, the green color changes to yellow and the spectrum shifts to a shorter wavelength. The reoxidized enzyme can be directly reduced by NADH2 to the green form without passing through a red-intermediate.
The authors wish to thank Dr. G. Sunagawa, the Director of this Laboratory, for his encouragement during the investigation.抄録全体を表示PDF形式でダウンロード (736K) -
JIRO KOYAMA, NOBUKO MIYAJIMA1966 年 60 巻 1 号 p. 27-36
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリーThe heterogeneity of guinea pig 7S γ-globulins and the rates of incorporation of C14-amino acids into electrophoretically dif-ferent 7S γ-globulin have been investigated.
1. Experiments in vivo showed that C14-amino acids were incorporated into electro-phoretically different fractions of guinea pig 7S r-globulins at different rates, indicating the presence of three different types of 7S γ-globulins which were referred to as slow 7S γ2-, fast 7S γ2- and 7S γ1-globulins.
2. The experiments in vitro with various tissues showed that the spleen, lymph nodes, bone marrow and lung incorporated actively C14-amino acids into both 7S γ1- and 7S γ2-globulins.
3. Furthermore, the expeviments in vitro revealed that C14-amino acids were incorpo-rated into different fractions of 7S γ2-globulin at different rates, depending on whether spleen, bone marrow, lymph nodes or lung was used.
4. From these results, it was suggested that guinea pig 7S γ1- and 7 S γ2-globulins were synthesized by different types of plasma cells and also that electrophoretically different 7S γ2-globulins were synthesized by different cells which were unevenly distributed in the various tissues.
5. Immunological examinations of guinea pig 7S γ-globulins for antigenicity were also performed and a result suggesting three sub-classes of 7S γ-globulin (slow 7S γ2-, fast 7S γ2- and 7S γ1-globulins) was obtained.
The present authors wish to express their sincere thanks to Prof. I. Yamashina for his encouragement during this work.抄録全体を表示PDF形式でダウンロード (1293K) -
III. Molecular Change of Hemoglobin during Spontaneous MetamorphosisKINZABURO HAMADA, YOSHIO SAKAI, KEIZO TSUSHIMA, RYOITI SHUKUYA1966 年 60 巻 1 号 p. 37-41
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリーThe circulating blood of Rana catesbeiana tadpoles which are undergoing metamor-phosis contain both tadpole and adult frog hemoglobin. The appearance of frog hemo-globin in the blood of animals during metamorphosis can be followed by determi-nation of the reactive sulfhydryl groups in the blood for these are only present in frog hemoglobin. The time required for the complete molecular change from tadpole hemoglobin to frog hemoglobin is approxi-mately seven to eight days.
The authors wish to express their thanks to Dr. K. Kaziro for his advice. This work was supported by a Grant for Scientific Research from the Ministry of Education of Japan.抄録全体を表示PDF形式でダウンロード (351K) -
IV. Post-mortem Breakdown of Phosphoinositides and Phosphatidic Acid, and P32-Incorporation into Phospholipids in Various States of the TissueKOHEI HAYASHI, YOSHIYUKI YAGIHARA, ISAMU NAKAMURA, SABURO YAMAZOE1966 年 60 巻 1 号 p. 42-51
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリーPost-mortem breakdown of P32-labelled phosphoinositides and phosphatidic acid in guinea-pig brain was studied utililyzing liquid oxygen for fixation. The incorpora-tion of P32-phosphate into phosphoinositides and phosphatidic acid in brain tissue in various states of the tissue was also studied. The results are summarized as follows; 1) Depletion of P32-labelled triphosphoinositide on thawing brain was extremely rapid, amounting to 63 per cent after 10 mintes and 91 per cent after 30 minutes at 30°C. The depletions of P32-labelled di-, monophospho-inositide and phosphatidic acid were 52, 14 and 39 per cent respectively, after 10 minutes under the same conditions. 2) The incorpora-tion of P32-phosphate into polyphosphoinosi-tides in brain slices was much more stimulated by the addition of sodium ion, than that into phosphatidic acid. The degree of stimulation of incorporation into monophosphoinositide was smaller than that into polyphosphoinositide. 3) P32-incorpora-tion into phosphoinositides and phosphatidic acid in brain tissue was changed conside-rably by changes in the state of the tissue i.e, in tissue in vivo, in slices or homogenates, and by changes in the incubation medium. 4) Polyphosphoinositides in concentrated HC1-chloroform-methanol (1:200:100) were quite stable for at least 18 hours at 4°C.抄録全体を表示PDF形式でダウンロード (665K) -
V. Occurrence of 4-O-Methylgalactose in Corbicula GlycolipidOSAMU ITASAKA1966 年 60 巻 1 号 p. 52-55
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリーThe nature of neutral sugar components of the corbicula glycolipid, GL-3, eluted with chloroform-methanol (1:1v/v) from the silicic acid column was investigated. A new sugar which has not been detected in glycolipids was found to be a component of the corbicula glycoliyid and was identified to be 4-O-methylgalactose by gas chromato-graphy.
The author acknowledges the valuable discussions and suggestions of Prof. T. Hori of this laboratory and Prof. T. Yamakawa, University of Tokyo, during the course of this investigation. The author also wishes to express his thanks to Prof. S. Hirase, Kyoto Technical University, for the gift of 2-O-, 4-O- and 6-O-methylgalactose.抄録全体を表示PDF形式でダウンロード (251K) -
LXXXII. Comparative Studies on the Bile Salts of Fishes by Thin Layer ChromatographyTAKESHI SASAKI1966 年 60 巻 1 号 p. 56-62
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリーThe bile salts of about 30 species of fishes were examined by thin layer chro-matography with the following results.
1. The bile salts of Elasmobranchii consist mainly of scymnol sulfate but also contain tauro-cholate and chimaerol sulfate as minor constituents.
2. Small amount of cholic acid, allo-cholic acid and an unknown bile acid having the same chromatographic behaviour as dihydroxycholanic acid are present in biles of members of the family Cyprinidae, the chief bile salts of which was shown to be 5α-cyprinol sulfate.
3. The main bile constituent of Mis-urgurnus anguillicadatus is 5α-cyprinol sulfate and a little amount of trihydroxycholanic acid conjugated with taurine is also present.
4. Parasilurus asolus bile contains 5α-cyprinol sulfate as a minor constituent in addition to tauro-cholate and tauro-cheno-deoxycholate.
5. Oncorhynchus rhodurus bile contains 3-keto-7α, 12α-dihydroxycholanic acid and cholic acid.
6. In two species of the Class Anguillida, 5β-cyprinol sulfate was shown to be a minor bile component as is the case of Conger myriaster.
7. A new bile acid, haemulcholic acid was isolated from the bile of Parapristipoma trilineatum.
The author wishes to thank Prof. T. Kazuno and Dr. T. Hoshita for their helpful suggestions throughout this work.抄録全体を表示PDF形式でダウンロード (402K) -
YASUYUKI OGURA, YUMIKO OHTA, TAKAO NAKAMURA1966 年 60 巻 1 号 p. 63-71
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリーThe pH-effect on the overall reaction rate of yeast L (+)-lactate dehydrogenase [EC 1. 1. 2. 3] was investigated in the pH-range from 5.0 to 9.0, with due precaution to avoid the occurrence of irreversible loss of activity.
When methylene blue, thionine, 2, 6-dichlorophenol-indophenol or cytochrome c was used as acceptor, VpHm/Vopm-pH relations gave sigmoid curves of the first order on both acidic and alkaline sides (VpHm: maximum velocity at a given pH; Vopm: maximum velo-city at the optimum pH). The pK-values were found to be 5.1 and 9.2 at 15°C, and 5.25 and 8.9 at 25°C, independently of the kind of acceptors used. The VpH/Vop-pH relations were also obtained as sigmoid curves of the first order, (VpH: velocity at a given pH; Vop: velocity at the optimum pH; both in the presence of a given, non-saturating, concen-tration of substrate). The VpH/Vop-pH curve on the acidic side shifted toward alkaline side with the decrease of the concentration of substrate added. The pK-values were 5.6 and 5.9 at 15°C in the presence of 1.05 and 0.105 mM L (+)-lactate, respectively. The pK-values obtained on the alkaline side were 9.2 at 15°C independently of the concentration of substrate added. In the case of ferricyanide used as acceptor, however, the pK-values on the acidic and alkaline sides were 6.0 and 9.2 at 15°C independently of the concentrationn of substrate.
The p(K'm/Km)-pH relations which were obtained on the acidic side by applying methylene blue, thionine or 2, 6-dichloro-phenol-indophenol as acceptor, were expressed by a formula, K'm/Km=(1+[H+]/K1)/(1+[H+]/K'1), where K1 and K'1 were assumed to be 10-6.0 and 10-5.1M respectively. From the data obtained at 25°C, the values of K1 and K'1 were calculated to be 10-6.15 and l0-5.25 M by applying Dixon's graphic method. When ferricyanide was used as acceptor, the p(K'm/Km)-values were almcst zero independently of the pH in the reaction media.
A reaction scheme was proposed to account for the various kinetic phenomena observed.
The authors wish to express their gratitudes to Prof. Hiroshi Tamiya for his valuable criticism in this work. We are indebted to Mr. T. Nagashima for his technical assistance. Thanks are also due to the Oriental Yeast Co. for the supply of baker's yeast.抄録全体を表示PDF形式でダウンロード (739K) -
V. Studies on Velocity Constants for Association and DissociationKINJI KAKIUCHI, TOSHIZO ISEMURA1966 年 60 巻 1 号 p. 72-76
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリーI. The velocity constant for dissociation, km, was determined from sedimentation velocity measurements by slightly modifying and directly applying the theoretical equa-tions, and was found to be 1×10-4 mole/liter-sec. The association velocity constant, kD, could be calculated from the association constant as K=2.2×109 (mole/liter)-2, (given by the sedimentation equilibrium measure-ments) and the dissociation velocity constant, and was found to be 2×105 mole/liter. sec.
2. In the previous papers, it was reported that the monomer cf B. α-amylase scarcely exists at concentrations above 0.4 percent protein. Using a synthetic boundary cell, a stable artificial boundary could be created by layering a solution of 0.504 per cent protein over the 1.07 per cent protein solution. The artificial boundary which was formed between the 0.504 ands 1.07 per cent solutions indicated the existance of the dimer alone, while the natural boundary which was somewhat skewed at the trailing edge was suggested to dissociate partly into monomer owing to decrease in the B. α-amylase con-centration to below 0.4 per cent.
The authors wish to express their gratitude to Dr. Shoichi Ikeda for valuable discussion and also their thanks to the Daiwa Kasei Co., Ltd., for supplying Bacillus sublilis α-amylase.抄録全体を表示PDF形式でダウンロード (1077K) -
YASUYUKI OGURA, TAKAO NAKAMURA1966 年 60 巻 1 号 p. 77-86
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリー1. When the reduced form of yeast L (+)-lactate dehydrogenase [EC 1. 1. 2. 3] was oxidized by molecular oxygen or ferricyanide, an absorption spectrum having maxima at 533 and 567.5 mμ appeared. It was confirmed that the enzyme having the absorption maxima at 533 and 567.5 mμ is in the true oxidized form containing trivalent iron atoms. The difference absorption spectrum at the steady state of reaction with ferricyanide as acceptor, which was followed by the “stopped flow” method, was in agreement with that of reduced minus oxidized form of the enzyme.
2. The rate of reduction of the protoheme moiety was measured by the “stopped flow” method in the presence of substrate and acceptor.
Except in the case of methylene blue used as acceptor, the kmaxred/2-values (kmaxred: apparent rate constant of the reduction of protoheme at saturating concentrations of the substrate) at pH 7.2 showed a close agreement with those of the maximum rate (Vm/e) obtained by the overall reaction kinetics. The value of kmaxred/2 was markedly larger than that of Vm/e when methylene blue was used as acceptor. It was inferred that, except in the case of methylene blue and under the experimental conditions used, the acceptors receive electrons mainly through the protoheme moiety, whereas methylene blue accepts electrons partially from protoheme and partially from the reduced flavin moiety.
At pH 5.2, kmaxred/2=Vm/e when ferricyanide was used as acceptor, while kmaxred/2<Vm/e in the cases of other acceptors. This fact was construed as indicating that ferricyanide molecules can accept electrons only from the reduced protoheme, while at pH 5.2 other acceptors (methylene blue, thionine et_??_.) accept electrons mainly from the reduced flavin moiety.
3. From the relationship beetween kmaxred and pH, the pK value at 25°C was found to be 6.1 independently of the kind of acceptors used. -values, which were obtained
4. The kmaxred-values, which were obtained at pH 5.2 and 7.2 using ferricyanide as acceptor, were both found to be independent of the concentration of the enzyme added. The data obtained at pH 5.2 seemed to be in favor of the assumption that the electron-transfer from flavin to protoheme be an intramolecular reaction process.
5. The reaction scheme proposed in the preceding paper (1) was shown to be also capable of explaining various experimental facts observed in the present study.
The authors wish to express their gratitude to Prof. H. Tamiya for his valuable criticism in this work. Thanks are also due to Oriental Yeast Co. for the supply of baker's yeast.抄録全体を表示PDF形式でダウンロード (769K) -
TOSHIO ONISHI, MUTUMI MURAMATU, TOSHIO SATO, SETSURO FUJII1966 年 60 巻 1 号 p. 87-89
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリーPDF形式でダウンロード (203K) -
TADASHI ASAHI, SHIGEHIDE MASAKI1966 年 60 巻 1 号 p. 90-92
発行日: 1966/07/25
公開日: 2008/11/18
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HIDEO INOUE, KUNIO HONJO, YOSHIRO TAKEDA1966 年 60 巻 1 号 p. 93-95
発行日: 1966/07/25
公開日: 2008/11/18
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MAKOTO SEIJI, TOSHIO YOSHIDA1966 年 60 巻 1 号 p. 96-98
発行日: 1966/07/25
公開日: 2008/11/18
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MICHIHIKO OGUCHI, TAKASHI HASHIMOTO, SHIGEKI MINAKAMI, HARUHISA YOSHIK ...1966 年 60 巻 1 号 p. 99-101
発行日: 1966/07/25
公開日: 2008/11/18
ジャーナル フリーPDF形式でダウンロード (1305K)
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