The Journal of Biochemistry
Online ISSN : 1756-2651
Print ISSN : 0021-924X
Volume 63, Issue 5
Displaying 1-19 of 19 articles from this issue
  • KEIICHI KUSAMA
    1968 Volume 63 Issue 5 Pages 561-565
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
    JOURNAL FREE ACCESS
    1. Tritiated uridine was degraded by the periodate and phenyl-hydrazine method to uracil without release of 3H from pyrimidine ring at the C5 position but, by treatment with 12 N perchloric acid, the C5-3H was completely eliminated.
    2. The release of C5-3H from uracil and uridine by boiling with 1 N acid was first order and the velocity constants were calculated. The C6-3H was very stable to this acid treatment.
    3. The distribution of 3H in commercial 5, 6-3H uracil and 5, 6-3H uridine, which were tritiated by catalytic exchange in 3H2O, was measured by bromination and by acid treatment. The percentage of III at C6 was 0.7-0.8% and 2-4 % of the total 3H bound in the pyrimidine ring of uracil and uridine, respectively.
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  • III. On the Binding of Chromomycin A3 with DNA and Physicochemical Properties of the Complex
    MIKIO KAMIYAMA
    1968 Volume 63 Issue 5 Pages 566-572
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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    1. A shift of the absorption maximum of chromomycin A3 at 406 mμ toward longer wavelengths was found in the presence of Mg++ or other bivalent cations such as Mn++, Co++ and Ni++. Both ultraviolet and visible absorption bands of chromomycin A3 were intensified by the addition of Mg++.
    2. Native and heat-denatured calf thymus DNA and yeast tRNA did not show any effect on the spectrum of the antibiotic both in the presence and in the absence of Mg++.
    3. In the presence of Mg++, the thermal transition curve of calf thymus DNA shifted toward higher temperatures on addition of the antibiotic.
    4. The Mg++ requirement for binding of chromomycin A3 with DNA was demonstrated by the fact that DNA precipitated by ethanol was yellow in the presence of Mg++ but white in its absence and that the absorption of the supernatant at 406 mμ decreased in the presence of Mg++ but not in its absence.
    5. The sugar moiety of chromomycin A3 was necessary for binding of the antibiotic with DNA, because chromomycinone did not co-precipitated with calf thymus DNA in the presence of Mg++.
    6. Chromomycin A3 co-sedimented with DNA only in the presence of Mg++ as examined by ultracentrifugation in a sucrose solution.
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  • I. Preparation and Properties
    EDAHIKO MURAKAMI
    1968 Volume 63 Issue 5 Pages 573-577
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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    1. Xanthurenic acid(XA) and insulin readly combine on simple mixing of their solutions.
    2. The complex shows a new absorption maximum at 327.5mμ and this maximum is the same in position as that of a chelate compound of XA, zinc and histidine.
    3. There are two moles of XA per dimer of insulin in the complex, so that the XA molecule probably combines with the imidazole group of a histidine residue of insulin via a zinc atom.
    4. The specificity of the binding of XA to protein was studied.
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  • II. Physiological Activities
    YAHITO KOTAKE, TOSHIKO SOTOKAWA, EDAHIKO MURAKAMI, AYAKO HISATAKE, MAS ...
    1968 Volume 63 Issue 5 Pages 578-581
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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    1. The activity of the xanthurenic acid-insulin complex on the blood sugar levels of rabbits is about 50% of that of native insulin.
    2. Formation of the complex also decreased the activities of insulin on the uptake of glucose by epididymal fat pads and by isolated diaphragms of rats.
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  • AKIRA OGAMO, YASUO SUZUKI, SEIICHI OKUI
    1968 Volume 63 Issue 5 Pages 582-590
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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    The properties of rat liver mitochondria treated with acetone con-taining various proportions of water were investigated. Acetone con-taining one percent of water extracted ubiquinone (UQ) quantitatively from mitochondria without causing any significant change in the con-tent of phospholipid. This preparation consumed oxygen with succinate as substrate on addition of cytochrome c alone, and UQ had little in-fluence on this activity. Thus, oxygen seemed to be consumed via a UQ independent pathway.
    A phosphate buffer extract (F-S) of mitochondria which were pre-treated with acetone containing one percent water inhibited the succinate-UQ reductase activity of the insoluble residue (R). From the effects of heat, gel filtration of Sephadex, and chymotrypsin [EC 3.4.4.5] treatment, the inhibitory factor in F-S seemed to be a protein. This protein factor was not readily solubilized unless mitochondria were pre-treated with aqueous acetone. A marked inhibitory effect of F-S was observed when it was allowed to come into contact with UQ in a high ionic strength medium before incubation with R. The mechanism of inhibition of succinate-UQ reductase by F-S was discussed.
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  • GENTARO OKADA, KAZUTOSI NISIZAWA, HIROSHI SUZUKI
    1968 Volume 63 Issue 5 Pages 591-607
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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    Three cellulase [EC 3.2.1.4, β-1, 4-glucan glucanohydrolase] com-ponents were obtained from Meicelase, a commercial product from Trichoderma virile. The purification was carried out through consecutive column chromatography. Each cellulase showed a single peak in ultra-centrifugal pattern, and the sedimentation coefficients were 3.54S, 3.75S and 3.62S, respectively. Their amino acid compositions resembled each other in respect of high contents of acidic amino acids, glycine, serine and threonine, and of low contents of basic amino acids and sulphur containing amino acids. They contained carbohydrates in an amount of 10-15%, which appears to consist of several neutral sugars and glucosamine.
    The optimal activities of the three cellulases were similarly found to be at pH 4.5-5.2, but their heat stabilities were different from each other. The three cellulases hydrolyzed, with different spectra, a series of substrates including several cello-oligosaccharides, CMC and various insoluble celluloses such as Avicel and absorbent cotton, but showed no activities toward cellobiose and aryl β-glucoside.
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  • I. Effect of Diazobenzene-p-sulfonate Modification on the Activation of Myosin A ATPase by Activators
    TATSUHISA YAMASHITA, IZUMI KABASAWA, TAKAMITSU SEKINE
    1968 Volume 63 Issue 5 Pages 608-616
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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    1. Coupling of myosin A with diazobenzene-p-sulfonic acid in-hibited the EDTA-ATPase activity without any essential change in activity or catalytic properties of Ca++-ATPase. The coupling also selectively diminished the allosteric action of other effectors, DNP and NEM.
    2. Ionic strength, pH and substrate or its analogues had little or no effect on azo coupling of myosin with DBS.
    3. It was concluded from spectrophotometric comparison of azo-myosin with azo amino acids and from other experimental results that the sulfhydryl residues in the active site of myosin had reacted with DBS.
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  • KAZUKO OSHIMA, IKUZO URITANI
    1968 Volume 63 Issue 5 Pages 617-625
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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    When 1-C14-acetyl-CoA was incubated with an enzyme preparation obtained from the non-infected tissue adjacent to the infected region of sweet potato with black rot, a C14-labeled β-hydroxy-β-methylglutaric acid-derivative was synthesized. This was concluded from the fact that the main ethyl acetate-extractable substance after alkaline hydrolysis of the reaction products proved to be β-hydroxy-β-methylglutaric acid, as demonstrated by paper radiochromatography, paper radioautography and constancy of specific radioactivity through repeated crystallizations.
    The slight enzymatic activities were detected in both healthy and cut tissues where furanoterpenoids did not occur. The activity was higher in the tissue after 2 days of infection than after 1 day in parallel with the accumulation of furanoterpenoids.
    The enzymatic synthesis was localized both in microsomal and supernatant fractions. No essential cofactors have been detected for the reaction. Some sulfhydryl-blocking reagents strongly inhibited the enzymatic activity.
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  • YOSHIAKI NAKAMARU
    1968 Volume 63 Issue 5 Pages 626-631
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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    1. The distribution of the strophanthin-sensitive ATPase (SS-ATPase) and the Mg+Ca-ATPase, and the effect of deoxycholate (DOC) on these enzymes activities, were investigated in several fractions of pig brain.
    2. The presence of the Mg+Ca-ATPase in the brain microsomal fraction was clearly shown. It was also shown that the Mg+Ca-ATPase was very similar to the SS-ATPase in its distribution pattern in several fractions of the brain homogenate and in its sensitivity to DOC.
    3. The microsomal fraction extracted with 0.1% DOC solution exhibited high Mg+Ca-ATPase activity and high SS-ATPase activity. For maximum activity of the strophanthin-insensitive ATPase, 3×10-5 M Ca was necessary in the presence of 2 × 10-3 M Mg, and 4 × 10-3 M Mg was necessary in the presence of 3×10-5 M Ca.
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  • YASUNORI NITTA, KEITARO HIROMI, SÔZABURO ONO
    1968 Volume 63 Issue 5 Pages 632-636
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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    1. Maltose was found to be hydrolyzed at pH 5.3 and 25°C by Taka-amylase A [EC 3.2.1.1] preparation which was crystallized three times and which was proved to be of single protein component by electrophoretic and ultra centrifugal analyses.
    2. The Michaelis constant Km for the hydrolysis of maltose was found to be numerically equal to the competitive inhibitor constant Ki of maltose for the hydrolysis of p-nitrophenyl α-maltoside catalyzed by Taka-amylase A.
    3. This Km value for maltose is one hundred times larger than the Km value for the hydrolysis of maltose catalyzed by Taka-maltase [EC3.2.1.20] or Taka-amylase B [EC 3.2.1.3] preparation.
    4. A treatment of Taka-maltase at pH 8.8 and 37°C for 2 hr decreases its specific activity towards maltose, but the same treatment of the crystalline Taka-amylase A preparation causes no loss of specific activity towards maltose. On the other hand, a treatment with 1% HgCl2 solution at pH 6.8 and room temperature for 24 hr for crystalline Taka-amylase A preparation decreases its specific activity towards maltose, but the same treatment for Taka-amylase B causes no loss of specific activity towards maltose.
    5. From the results, it was concluded that the observed hydrolysis of maltose was caused by the action of Taka-amylase A itself and not by Taka-maltase or Taka-amylase B which might be contaminated in the crystalline preparation of Taka-amylase A.
    6. The Michaelis constant Km for the hydrolysis of maltose catalyzed by Taka-amylase A was determined to be 9.6 ×10-2 M and its breakdown rate constant k+2 of ES complex was determined to be 0.019 sec-1.
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  • V. Purification of the Phenylalanine, Lysine and Histidine Transfer Ribonucleic Acids
    MASAZUMI MIYAZAKI, SHOSUKE TAKEMURA
    1968 Volume 63 Issue 5 Pages 637-648
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
    JOURNAL FREE ACCESS
    By a series of chromatography on a DEAE-Sephadex column with the ammonium sulfate, phosphate and borate systems, the two phenyl-alanine and one histidine tRNA's were highly purified. The phenyl-alanine and lysine tRNA's were respectively resolved into three com-ponents, while the histidine-acceptor activity did not display the hetero-geneity.
    Distribution of oligonucleotide fragments produced on pancreatic RNase I [EC 2.7.7.16] digestion of the purified preparations was quite characteristic of the amino acid specific tRNA's and, on the other hand, quite similar among the multiple components specific for the same amino acid. Preliminary results on the sequence analyses of the phenyl-alanine tRNA I have revealed several important sequences. Especially the following fragments should be noted: 5'-terminal, pGpCp, 3'-termial, CpApCpCpA, common sequence, (Tp, Ψp, Cp)Gp, and large fragment, Ap2'OMeCpUp[(2'OMeGpAp, Ap, X)Ψp, Cp, ApUp]Gp, and so on. The difference between the phenylalanine tRNA's I and III had not yet been found in nucleotide sequence examined. On the other hand, certain different sequences, together with the similarity, were noted between Torula and Baker's yeast.
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  • MASACHIKA IRIE
    1968 Volume 63 Issue 5 Pages 649-653
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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    1. Kinetic parameters, Km and Vmax, of RNase Tl [EC 2.7.7.26] were obtained usineg dinucleoside phosphate, GpX (GpA, GpC, GpG and GpU), as substrate at pH 7.5 and 5.0. Km values of four GpX's were similar order of magnitude, but Vmax values of them were decreas-ing in the following order; GpC>GpG>GpA>GpU.
    2. From the pKm-pH profile of RNase Tl at pH from 5.0 to 2.5 using GpC as substrate, pKe value of 3.5 was obtained.
    3. From the pKi-pH profile of RNase Tl using G-2'-p as competitive inhibitor, the pKe value of 3.4 was also obtained.
    4. Based on the values obtained above, the possibility of the presence of a carboxyl residue in the active site of the enzyme was discussed.
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  • KIYOSHI IKEDA, KOZO HAMAGUCHI, SHUNSUKE MIGITA
    1968 Volume 63 Issue 5 Pages 654-660
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
    JOURNAL FREE ACCESS
    The circular dichroism (CD) of ten individual samples of Bence-Jones proteins (with five each of type K and type L), and a normal human immunoglobulin G and a myeloma immunoglobulin G of type K in 0.15m KCl was investigated over the wavelength range from 200 to 325mμ. All the CD spectra of these proteins had a negative ma-ximum at around 217mμ which is characteristic of the β structure. The negative ellipticity at this wavelength of the type K Bence Jones proteins was generally smaller than that of the type L Bence Jones proteins except in the case of one K and one L Bence Jones proteins. In the 220 to 240mμ region, three type K Bence Jones proteins exhibited a positive CD band at 228 or 234 mμ, while none of the type L Bence-Jones proteins showed any corresponding extremum. In a still longer wavelength region, the CD spectra of type K Bence Jones proteins had a small but distinct positive maximum between 292 and 298mμ, whereas the type L Bence Jones proteins showed a positive maximum between 300 and 310 mu. Only one of the five type L Bence Jones proteins gave a negative maximum at 310mμ. The CD spectra of a normal and a myeloma immunoglobulin G were very similar to those of Bence Jones proteins. One type K myeloma protein studied here gave a CD spectrum similar to those of type K Bence Jones proteins.
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  • I. Effect of Purine Nucleotides on Inosine-5'-phosphate Dehydrogenase, Xanthosine-5'-phosphate Aminase and Adenylosuccinate Lyase of Bacillus subtilis
    KENJI ISHII, ISAMU SHIIO
    1968 Volume 63 Issue 5 Pages 661-669
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
    JOURNAL FREE ACCESS
    The effects of purine and pyrimidine nucleotides and of the nucleosides on partially purified IMP dehydrogenase [EC 1.2.1.14], XMP aminase [EC 6.3.5.2] and succino-AMP lyase [EC 4.3.2.2] activities of Bacillus subtilis M strain of derepressed cells were studied. IMP dehydrogenase was inhibited strongly by GMP, the end product, and by XMP, the direct product of the reaction under dilute con-centrations of IMP and KC1. When the concentrations of IMP and KC1 were both high, ATP also was a strong inhibitor. The fact that Lineweaver-Burk plot against IMP curved upward in the presence of GMP and XMP and was linear in the absence indicated the enzyme was allosteric. XMP aminase utilized i-glutamine more effectively than ammonium sulfate as the amino donor and its activity was inhibited slightly by several ribonucleotides including GDP, GMP, adenosine, UDP, IDP, AMP and ITP. Succino-AMP lyase activity with succino-AMP as the substrate was inhibited competitively by the product, AMP, and its derivative, ATP.
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  • MAKOTO SEIJI, TOSHIO YOSHIDA
    1968 Volume 63 Issue 5 Pages 670-674
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
    JOURNAL FREE ACCESS
    To study the state of tyrosinase [EC 1. 10. 3. 1] in the smooth sur-faced vesicles, the smooth surfaced membrane fraction isolated from Harding-Passey mouse melanoma was digested by Naja naja venom phospholipase A [EC 3. 1. 1. 4]. The digest was centrifuged and separated into two fractions: the supernatant and the precipitate. The amounts of protein and phospholipid and various enzyme activities including tyrosinase in each were determined. The effect of digestion on tyros-inase was different from its effects on other enzymes. The amount of tyrosinase activity solubilized was proportional to the amount of protein in the supernatant and the specific activities in the supernatant and sediment were almost the same. Furthermore, most of the tyrosinase activity remained in the residue of digested membranes. It is con-cluded from these results, that tyrosinase in the smooth surfaced vesicles is tightly bound to the protein moiety in the membranous structure as a constitutional element of the membrane.
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  • SUSUMU KURIOKA
    1968 Volume 63 Issue 5 Pages 675-677
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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  • SUSUMU KURIOKA
    1968 Volume 63 Issue 5 Pages 678-680
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
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  • SHOJI MIZUSHIMA, MASAMITSU ITO
    1968 Volume 63 Issue 5 Pages 681-683
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
    JOURNAL FREE ACCESS
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  • MITSUO NAKAGAWA, MITSURU UCHIYAMA
    1968 Volume 63 Issue 5 Pages 684-687
    Published: May 25, 1968
    Released on J-STAGE: November 18, 2008
    JOURNAL FREE ACCESS
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