巻号一覧
64 巻, 4 号
選択された号の論文の19件中1～19を表示しています
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II. Separation of Exo- and Endopeptidases of PronaseYOSHIKO NARAHASHI, KUNITOYO SHIBUYA, MASAYA YANAGITA1968 年 64 巻 4 号 p. 427-437
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリーBy column chromatography with CM-cellulose, DEAE-Sephadex and Amberlite CG-50 resin, eleven proteolytic enzymes were fractionated from Pronase. They were four neutral proteinases, three alkaline proteinases, three aminopeptidases and carboxypeptidase. Chromatography of Pronase solution on CM-cellulose resolved enzymes into four broad peaks, peaks I, II, III and IV. Peaks I and II were mainly composed of neutral proteinases and aminopeptidases, peak III contained alkaline proteinases and carboxypeptidase and the last peak was alkaline proteinase. Among these enzymes, only the alkaline proteinase present in peak III possessed high activity towards N-benzoyl-L-arginine ethyl ester, N-α-tosyl-L-arginine methyl ester and N-benzoyl-L-arginine-p-nitroanilide. Further separation of aminopeptidase from neutral proteinase contaminated was accomplished by DEAE-Sephadex chromatography. On the other hand, carboxypeptidase was isolated by chromatography on Amberlite CG-50 resin from peak III. The neutral proteinases were inhibited by EDTA, whereas alkaline proteinases were inhibited by DEP but not by EDTA.抄録全体を表示PDF形式でダウンロード (1360K) -
SATOSHI NAKAMURA, YASUYUKI OGURA1968 年 64 巻 4 号 p. 439-447
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリー1. The reaction catalyzed by the glucose oxidase [EC 1. 1. 3. 4] from Aspergillus niger was markedly inhibited by Ag+, Hg++, Cu++, PCMB and PMA, but not by non-metallic SH-reagents, such as NEM, IA and IAA. No sulfhydryl group was detected in the enzyme protein by spectrophotometric titration with PCMB and NEM. Characteristic difference spectra due to the absorption by the FAD moiety of the enzyme were obtained on the addition of Ag+, Hg++, Cu++, PCMB and PMA to enzyme solutions while no spectral change was observed on the addition of other metal ions and NEM, which did not inhibit the enzyme reaction.
2. From the studies by overall reaction kinetics and the “flow method”, it was confirmed that Ag+ and Hg++ ions inhibit the oxidation of the reduced FAD moiety, competing with molecular oxygen used as a hydrogen acceptor. While Ag+ ion did not inhibit the reduction of the FAD moiety, Hg++ ion more or less inhibited the reduction described above. It was suggested that there is an interaction, probably by steric hindrance, between the substrate and the Hg++ ion for binding to the enzyme molecule. From the kinetic data obtained in the presence of both Ag+ and Hg++ ions, it was concluded that the binding sites for these two kinds of metal ions on the enzyme molecule are different.抄録全体を表示PDF形式でダウンロード (587K) -
KATSUHIDE YUTANI, ASAKO YUTANI, AKIRA IMANISHI, TOSHIZO ISEMURA1968 年 64 巻 4 号 p. 449-455
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリー1. Renaturation process of the reduced random coil form of lysozyme [EC 3. 2. 1. 17] was studied by various physicochemical measurements such as difference spectrum, optical rotatory dispersion and circular dichroism. The data were related to the extent of reoxidation of sulfhydryl groups and recovery of the enzymatic activity.
2. About seventy per cent of the original helical content of the native enzyme was reformed, immediately after the dilution of the reduced lysozyme in 8 M urea with a Tris-Cl buffer solution, pH 8. 0. In this partial reformation of the internal fold, one or two of six tryptophan residues of lysozyme was involved.
3. During reoxidation, one more tryptophan residue per enzyme molecule was buried in the internal fold, and enzymatic activity was gradually recovered. This reformation of the original conformation of the enzyme may proceed through the interchange of disulfide bonds as well as that of noncovalent bonds.抄録全体を表示PDF形式でダウンロード (461K) -
GOTARO TODA1968 年 64 巻 4 号 p. 457-464
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリー1. When Na-K ATPase [EC 3. 6. 1. 3] preparation from guinea pig kidney cortex was preincubated with BeCl2 in the presence of Mg++, the activity was inhibited.
2. For beryllium inhibition of Na-K ATPase, Mg++ was necessary and was substituted by Mn++, but not by Ca++
3. Beryllium chloride concentration required for 50% of Na-K ATPase activity was 1.8×10-6M.
4. In the presence of Mg++, K+ increased the rate of inhibition and was substituted by NH4+ or Rb+, but not by Li+.
5. Sodium ion not only protected the enzyme from the inhibition in the presence of K+ and Mg++, but also reversed the inhibition.
6. Protective effect of Na+ was remarkable in the absence of K+ or presence of its low concentration.
7. Ouabain did not affect the rate of inhibition in the presence of K+ and Mg++.抄録全体を表示PDF形式でダウンロード (572K) -
I. Preparation and Physiological FunctionSETSURO EBASHI, AYAKO KODAMA, FUMIKO EBASHI1968 年 64 巻 4 号 p. 465-477
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリー1. A method for isolation of troponin from native tropomyosin was described.
2. Troponin in combination with tropomyosin restored the whole activity of native tropomyosin in sensitizing the interaction of myosin and actin to Ca ion.
3. Troponin was found to bind nearly 4 moles of Ca per 105g, of which most were exchangeable. The result of the experiment to determine the binding constant of these Ca binding sites was explained by assuming that half of the binding sites possessed a binding constant of 1.3×106 M-1 and the remaining half 5×104 M-1.
4. The amount of exchangeable Ca in the contractile system was mainly accounted for by the Ca-binding capacity of troponin, which was not influenced by other contractile proteins or ATP.
5. Cardiac troponin showed a much higher affinity for Sr ion than skeletal troponin. The ratio of the former affinity to the latter was in good agreement with the ratio of the sensitivity to Sr ion of a reconstituted contractile system containing cardiac troponin to that containing skeletal troponin. Based on these findings and the results described above, it was concluded that the sensitivity of a contractile system to Ca ion is solely dependent upon the affinity for Ca ion of the troponin molecule present.
6. The mechanism of troponin regulation of the interaction of actin and myosin was discussed.抄録全体を表示PDF形式でダウンロード (988K) -
YOSHIO UENO, MASAKATSU HOSOYA, YUKO MORITA, IKUKO UENO, TAKASHI TATSUN ...1968 年 64 巻 4 号 p. 479-485
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリーIn order to find out a bioassay method of the toxic metabolites of Fusarium nivale, an investigation was made with following results;
1. Protein synthetic reaction of the rabbit reticulocyte was applicable to the fractionation of the toxic principle from Fusarium nivalegrowing rice.
2. The isolated toxin, Nivalenol, inhibited the protein synthesis at the ribosomal level of the cell.抄録全体を表示PDF形式でダウンロード (483K) -
I. Preparation and Properties of Tritium Labelled SphingoglycolipidsYOUSUKE SEYAMA, TAMIO YAMAKAWA, TOMOYOSHI KOMAI1968 年 64 巻 4 号 p. 487-493
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリーThe possibility of applying isotope dilution analysis in determination of individual sphingoglycolipids was examined. Tritium labelled sphingoglycolipids were prepared by catalytic reduction with tritium. The labelled compounds were clearly separated from unlabelled compounds on silicic acid column and silica gel thin layer chromatographies. This was found to be due to the disappearance of double bonds in the sphingosine bases and fatty acids during the reduction procedure. There fore, sphingoglycolipids should be determined by isotope dilution analysis using samples in which the double bonds have first been saturated by catalytic reduction.抄録全体を表示PDF形式でダウンロード (444K) -
FUJINORI NAKADA, REIJI OSHIO, SETSUO SASAKI, HAJIME YAMASAKI, NOBUO YA ...1968 年 64 巻 4 号 p. 495-501
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリー3β-Acetoxychol-5-enoic-24-14C acid was administered to a rat with a bile fistula. Bile samples were collected, and the following natural and unnatural bile acids were identified as metabolites: 3α- and 3β-hydroxy-cholanoic acids of the 5β- and 5α-series; 3_??_, 6_??_-dihydroxycholanoic acids of the 5β- and 5α-series which were identified as 3, 6-dioxo-5β-cholanoic and 3, 6-dioxo-5α-cholanoic acids after oxidation; 3α, 7α-, 3α, 7β- and possibly 3β, 7ξ-dihydroxy-5β-cholanoic acids. From this result a tentative pathway leading to those metabolites is presented.抄録全体を表示PDF形式でダウンロード (402K) -
KOZO HAMAGUCHI, KIYOSHI IKEDA, CHEN-YUAN LEE1968 年 64 巻 4 号 p. 503-506
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリーThe optical rotatory dispersion (ORD) and circular dichroism (CD) of the neurotoxins (α- and β-Bungarotoxin) isolated from the venom of Bungarus multicinctus were measured over the wavelength range of 195 to 300mμ. The ORD curve of α-Bungarotoxin had a positive peak at 207mμ and a negative trough at 220mμ. The CD spectrum of this protein showed a negative maximum at 216mμ, a positive maximum at 232 mμ and a shoulder at around 240mμ. Thus, it is suggested that α-Bungarotoxin molecule contains β-structure. On the other hand, the ORD curve of β-Bungarotoxin had a positive maximum at 198mμ and a negative trough at 232mμ. The CD spectrum had a negative maximum at 210mμ and a shoulder at 222mμ. Therefore, β-Bungarotoxin molecule contains α-helical structure. The effects of pH and 2-chloroethanol on the ORD and CD curves of these toxins were also studied.抄録全体を表示PDF形式でダウンロード (267K) -
XVI. Naphthoquinones with two Sulfonic Groups as a Reagent for Discrimination of Amino Groups in ProteinsAYAKO MATSUSHIMA, KATSUKIYO SAKURAI, MASAO NOMOTO, YUJI INADA, KAZUO S ...1968 年 64 巻 4 号 p. 507-514
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリーThe reagents so far used for partial modification of amino groups in proteins give rise to modified proteins which are unstable in aqueous solutions because of the lowering of solubility by the modification. In these circumstances, β-naphthoquinone-4, 5-, -4, 6- and -4, 7-disulfonic acids were examined as reagents for modification of amino groups and applied to several proteins, and the following results were obtained. The 4, 6- and 4, 7-disulfonic derivatives possess a moderate reactivity with amino groups near neutral pH (optimum pH between 8.4 and 9.1) and at room temperature, which is suitable for discrimination of different states of amino groups in proteins, and yet the proteins modified with these reagents are stable in solution. On the other hand, the 4, 5- disulfonic derivative is not reactive with amino groups under the same conditions. The degree of reaction can be determined simply by difference spectrophotometry at 480mμ between the sample and blank solutions. Approximately 2 of the total 3 amino groups in the insulin molecule and 9 of the 11 amino groups in the ribonuclease [EC 2. 7. 7. 16] molecule are reactive with the 4, 6- or 4, 7-disulfonic derivatives. About 6 of the total 7 amino groups in the lysozyme [EC 3. 2. 1. 17] molecule are modified with these reagents and the lytic activity is decreased by the modification to less than 3% of the original activity. In the case of trypsin [EC 3. 4. 4. 4], 25-40% of the activity is retained even after modification of almost all of the amino groups. Treatments of two samples of α-chymotrypsin [EC 3. 4. 4. 5] purchased from different companies with the reagents gave different results ; 14 and 17 amino groups per molecule are modified with the reagents and the activity drops to 60% and less than 6% on the modifications, respectively. At least one of the amino groups was inferred from these data to participate in the enzyme activity.抄録全体を表示PDF形式でダウンロード (588K) -
TOSHIO ASAKURA, HARUHISA YOSHIKAWA, KAZUTOMO IMAHORI1968 年 64 巻 4 号 p. 515-520
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリー1. Etiohemoglobin and dimethylprotohemoglobin were prepared from corresponding hemes and apohemoglobin. These two artificial hemoglobins showed higher affinity for oxygen and ethylisocyanide (EIC) than natural hemoglobin. The “n” value in Hill's equation was calculated as 1. 3 from EIC dissociation curves, indicating decreased heme heme interaction.
2. The half-denaturation temperature of etiohemoglobin and dimethylprotohemoglobin as well as of apohemoglobin was 41°, in contrast to 61° in protohemoglobin, reconstituted protohemoglobin and protoporphyrin-globin.
3. Sedimentation constants of etiohemoglobin and dimethylproto-hemoglobin were the same as that of apohemoglobin.
4. The rotatory dispersions of etiohemoglobin and dimethylproto-hemoglobin indicated that the helical contents of these proteins were lower than that of protohemoglobin.抄録全体を表示PDF形式でダウンロード (404K) -
TAKASHI MURAMATSU1968 年 64 巻 4 号 p. 521-531
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリーThe liver of a gastropod, Turbo cornutus contained α-N-acetylglucos-aminidase, β-N-acetylglucosaminidase [EC 3. 2. 1. 30], α-N-acetylgalactos-aminidase and β-N-acetylgalactosaminidase. The optimum pH value of the all four enzymes was near 4.0. Sephadex G-200 column chromato-graphy and heat treatment showed that α-N-acetylglucosaminidase, β-N-acetylglucosaminidase and α-N-acetylgalactosaminidase were different enzymes. β-N-Acetylglucosaminidase and β-N-acetylgalactosaminidase were not separated from each other by the above procedures, and the identity of the two enzymes was demonstrated kinetically.
α-N-Acetylglucosaminidase from Turbo cornutus was purified 76-fold with a recovery of 24%. The purified enzyme preparation contained small amounts of β-N-acetylglucosaminidase, β-glucosidase [EC 3. 2. 1. 21] and α-galactosidase [EC 3. 2. 1. 22], but was practically free from other glycosidases. The enzyme was activated by sodium chloride and inhibited by PCMB.
β-N-Acetylglucosaminidase from Turbo cornutus was purified 16-fold. The purified enzyme was practically free from all other glycosidase ac-tivities except that of β-N-acetylgalactosaminidase. The enzyme was activated by sodium chloride and inhibited by acetate. It released 2.7 moles of N-acetylglucosamine per mole of ovomucoid, without any release of hexose.抄録全体を表示PDF形式でダウンロード (711K) -
IX. Enzymatic Hydrolysis of Ceramide 2-Aminoethylphosphonate and SphingoethanolamineTARO HORI, IKUKO ARAKAWA, MUTSUMI SUGITA, OSAMU ITASAKA1968 年 64 巻 4 号 p. 533-536
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリーThe hydrolysis by Clostridium perfringens (welchii) phospholipase c [EC 3. 1. 4. 3] of ceramide 2-aminoethylphosphonate (CAEP) and sphingo-ethanolamine (Sph. EA), which are analogues of sphingomyelin, was studied. 2-Aminoethylphosphonic acid (ciliatine) and ceramide produced from CAEP by the phospholipase c, together with phosphorylethanol-amine and ceramide from Sph. EA were isolated in crystalline form, and identified.抄録全体を表示PDF形式でダウンロード (216K) -
III. Chemical Modification of D-Amino-Acid Oxidase Caused by GlyoxalAKIRA KOTAKI, MINORU HARADA, KUNIO YAGI1968 年 64 巻 4 号 p. 537-548
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリーChemical modification of arginine residues of D-amino-acid oxidase [EC 1. 4. 3. 3, D-amino-acid: oxygen oxidoreductase (deaminating)] was brought about at pH 8. 1 and at room temperature using glyoxal as their blocking agent, in the absence or presence of benzoate which forms a 1:1 complex with the enzyme on the molecular basis of 50, 000. The free enzyme was inactivated partially in proportion to the extent of modification of its arginine residues, and finally completely when reactive 18 out of 28 residues per molecule were modified. On the other hand, considerable resistance against inactivation was found when the enzyme previously formed a complex with benzoate; about 50% of the enzyme activity still remained even after the modification of reactive 17 residues per molecule.
The enzyme inactivated in the absence of benzoate completely lost the ability to form a complex with benzoate, whereas in the presence of benzoate the enzymes whose activities were lowered to about 50% by glyoxal still kept the full ability of complexing with benzoate. The yellow color of the modified enzyme preparation was completely bleached on addition of D-alanine under anaerobic conditions. No significant difference was found between the apparent Km value of the intact enzyme and that of the 50%-active enzyme. The apparent Vmax value of this modified enzyme, however, was lowered to about a half that of the intact enzyme.
Optical activity of the flavin chromophore of the modified enzyme was found to be essentially identical with that of the intact enzyme.
These results indicate the complete protection of the binding sites of the enzyme by complexing with benzoate and, therefore, strongly suggest the presence of a side group in the protein molecule, most probably one specific arginine residue that contributes to the enzyme action through its binding with the carboxylate group of the substrate.抄録全体を表示PDF形式でダウンロード (1433K) -
SHUZO YAMAGATA, RYO SATO1968 年 64 巻 4 号 p. 549-556
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリーA c-type cytochrome was highly purified from nuclei of calf thymus gland by deoxyribonuclease digestion, salt extraction, ammonium sulfate fractionation, and Amberlite CG-50 column chromatography. Cytochrome c was also purified from calf thymus mitochondria by the same procedure. The two cytochrome c preparations were found to be indistinguishable from each other in spectral properties, redox potential, electrophoretic behavior, reactivities, amino acid composition and peptide map pattern. It was concluded that the thymocyte contains the same species of cytochrome c in both the nucleus and mitochondria.抄録全体を表示PDF形式でダウンロード (550K) -
KUNIHIKO SAITO, MASAMI GAMO1968 年 64 巻 4 号 p. 557-559
発行日: 1968/10/25
公開日: 2008/06/30
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SHIGEKI MINAKAMI, E. N. MOKHOVA1968 年 64 巻 4 号 p. 561-562
発行日: 1968/10/25
公開日: 2008/06/30
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SHIGEHIRO HIRANO, HIROKO HAYASHI, TAKESHI TERABAYASHI, KONOSHIN ONODER ...1968 年 64 巻 4 号 p. 563-565
発行日: 1968/10/25
公開日: 2008/06/30
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KUNIO YAGI, TAKAYUKI OZAWA, NOBUKO OHISHI1968 年 64 巻 4 号 p. 567-569
発行日: 1968/10/25
公開日: 2008/06/30
ジャーナル フリーPDF形式でダウンロード (835K)
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