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Volume 56 (1964)
- Issue 6 Pages 499-
- Issue 5 Pages 381-
- Issue 4 Pages 299-
- Issue 3 Pages 201-
- Issue 2 Pages 113-
- Issue 1 Pages 1-
Volume 56, Issue 4
Displaying 1-15 of 15 articles from this issue
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AKIRA KOTAKI, KAZUO SATAKE1964 Volume 56 Issue 4 Pages 299-307
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Most of TNP-amino acids could be hydrolyzed with N sodium hydroxide at room temperature and gave picric acid and the 'corresponding free amino acids quantitatively. However, TNP-glycine, -serine and -threonine were decomposed into unidentified colored substances, without producing an appreciable amount of picric acid and free amino acids.
2. TNP-Amino acids were rather stable in acid. For example, no appreciable amount of TNP-valine was hydrolyzed with 6N hydrochloric acid, and more than 80 per cent remained intact even with 12N acid, after the treatment for 72 hours at 37°C. TNP-α-NH linkage of TNP-amino acids and -pep-tides, however, was cleaved quantitatively into picric acid and original free amino acids by heating with 6N hydrochloric acid at 110°C for 8 to 10 hours. TNP-ω-NH linkages were rather resistant to the conditions, and more, than 60 per cent were capable of remaining intact even after the prolonged heating for 24 hours in 6N hydrochloric acid. Extreme stability of TNP-amines in acid was also observed.
3. Analysis of TNP-peptides produced by the partial acid or enzymatic hydrolysis of TNP-protein was described. Amino acid contents of these TNP-peptides were able to be represented as the relative ratios to the contents of picric acid, liberated.View full abstractDownload PDF (579K) -
III. Induced synthesis of apo-tryptophan peroxidase molecule and protohemin IXKATSURO KOIKE, SEIICHI OKUI1964 Volume 56 Issue 4 Pages 308-313
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. By the DEAE-cellulose chromatographic procedure, the apo-tryptophan peroxidase activity localized in the fraction C of induced rat liver preparation (eluted with 0.2M po-tassium phosphate buffer containing 10-3M L-tryptophan, pH 7.0), and the protein content was about 6-fold of the normal liver preparation. C14-Glycine incorporation into apotryptophan peroxidase and tryptophan peroxidase activity in the presence of added hematin were about 6-fold increase as compared with normal rat liver preparation. There is a partly similar mechanism to the induced enzyme protein synthesis in microorganism.
2. The induced increment of tryptophan peroxidase activity by L-tryptophan in animal liver is due to the conversion of the newly synthesized heme- free apo-tryptophan peroxidase to an active holoenzyme with the incre-ased protohemin IX which showed about 3-fold elevation of its synthesis, and which is an activator or a prosthetic group of tryptophan peroxidase.View full abstractDownload PDF (432K) -
YASUHIRO KAWANISHI, KOICHI IWAI, TOSHIO ANDO1964 Volume 56 Issue 4 Pages 314-324
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Some improvements made on the fractional extraction procedure (see Scheme 1) of dinitrophenylated hydrazinolyzate of peptide or protein, could cause a quantitative separation of DNP-compounds, especially diDNP-derivatives, with free carboxyl group derived from the C-terminal.
2. Aspartic acid β-hydrazide and glutamic acid γ-hydrazide, both owing to the C-terminal residues, and aspartic or glutamic acid α-hydrazide owing to the intrachain residues were stabilized as the diDNP-derivatives, and successfully separated and identified on the two-dimensional paper chromatogram.
3. Catalytic hydrazinolysis according to Bradbury's method (heating at 60°C in hydrazine containing M hydrazine sulfate as catalyst) gave a high and reproducible yield of ornithine, aspartic acid β-hydrazide or glutamic acid γ-hydrazide as the diDNP-derivative from the C-terminal arginine or asparagine residue, or from glutathione.
4. Thus, combining the catalytic hydrazinolysis with the modified extraction and the paper chromatography, it became possible to determine quantitatively the C-terminal argi-nine and asparagine (and glutamine) residues of proteins in amounts necessary for the paper-chromatographic identification. The method was confirmed to be effective on such proteins as protamines and insulins from several sources, egg-white lysozyme, and carboxypeptidase-A.
The authors are indebted to Dr. K. Narita of Osaka University, Dr. K. Ohno of Ajinomoto Co., and Dr. K. Kusama of Shizuoka University for their helpful advices to the experiments.View full abstractDownload PDF (713K) -
II. Molecular Weight and Some Physicochemical Properties of Crystalline Ricin DMASATSUNE ISHIGURO, TAKAO TAKAHASHI, KATSUYA HAYASHI, MASARU FUNATSU1964 Volume 56 Issue 4 Pages 325-327
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSInvestigation of molecular weight and some physicochemical properties of ricin D was carried out. Sedimentation constant, s20, was calculated to be 4.64×10-13 in acetate buffer solution (μ=0.1) of pH 5.0. Molecular weight of ricin D was measured by Archibald's method and calculated to be 6.00×104. Electrophoretic mobilities were measured at pH values 3.6, 5.0, 6.0, 8.0, and 9.0 respectively and the isoelectric point of ricin D was found to be pH 5.9. Ultraviolet absorption spectrum of ricin D was consistent with that of a common protein.View full abstractDownload PDF (172K) -
III. Dinitrophenylation Studies on the Purified Yeast RNAHIDEO GOTO1964 Volume 56 Issue 4 Pages 328-334
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Commercial yeast RNA was extremely highly purified. The DNP-derivatives of such RNA (DNP-RNA) liberated several kinds of DNP-amino acids by alkali treatment and also by acid hydrolysis of the remainder of the alkali treatment. It seems likely that at least some parts of the amino acids liberated have originally been bound to the RNA in a more stable form, in which the NH2-groups of the amino acids are not involved.
2. Ultraviolet absorption spectra and infrared absorption spectra were compared between RNA and DNP-RNA before and after alkali treatment. Quantitative deter-mination of DNP-group introduced into RNA molecule was also made. These data indicated that practically no dinitrophenylation had occurred in purines and pyrimidines.
3. A peptide fraction was separated as DNP-derivatives from DNP-RNA or from a mixture obtained after dinitrophenylation of the alkali digest of RNA. Acid hydrolysis, of the peptide fraction yielded DNP-derivatives of histidine, serine, alanine and aspertic acid, indicating the occurrence in the RNA of not less than four different peptides having these amino acids as terminus.
The author wishes to express his sincere gratitude to Prof. S. Akashi for his guidance and counsel during the course of the work, and to Dr. T. Murachi for helpful advice in the preparation of the manuscript.View full abstractDownload PDF (1176K) -
TOMOJI SUZUKI, KYOZO HAYASHI, KAZUO FUJIKAWA, KYOZO TSUKAMOTO1964 Volume 56 Issue 4 Pages 335-343
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSIn order to determine the total structure, polymyxin B1 has been enzymatically hydro-lyzed with Subtilopeptidase A and the result-ing hydrolysate was fractionated by counter-current distribution. Four peptides have been isolated and their amino acid sequences were determined, and the peptides, MOA→(α)L-DAB→L-Thr→(α)L-DAB and cyclo-(γ)L-DAB→(α)L-DAB→D-Phe→L-Leu→(α)L-DAB→(α)L-DAB→L-Thr→, which were essential to eluci-date the full structure of polymyxin B1, were obtained. It was thus determined that the side chain is linked to the α-amino group of an α, γ-diaminobutyric acid residue in the cyclic peptide portion and the γ-amino group of this residue is involved in the ring forma-tion, and that the amino acids present in polymyxin B1 are contained as the L-configu-ration except phenylalanine. The structure of polymyxin B1 was thus elucidated to be MOA→(α)L-DAB→L-Thr→(α)L-DAB→cyclo-(γ)L-DAB→(α)L-DAB→D-Phe→L-Leu →(α) L-DAB→(α)L-DAB→L-Thr→.
The authors are extremely grateful to Dr. Craig of Rockefeller Institute and to the Pfizer & Co., Inc. for providing us polymyxin B sulfate. The authors are indebted to Research Laboratory of Shionogi & Co., Ltd. for the measurement of the rotatory dis-person and to Dr. K. Machida of Kyoto University for the measurement of the infrared spectra and also wish to express their thanks to Dr. T. Kawano and Miss M. Tsuchiya of our laboratory for their assist-ances.
Note added in proof.
In the personal communication received from Dr. K. Vogler of the Hoffmann-La Roche & Co. after submission of this paper, he informed us that he and his co-workers, according to our communication, have synthesized the compound which has the same structure (7α, all L-α, γ-diaminobutyric acid) as proposed by authors in this paper and that from the results of studies of the thin-layer chromatography, amino acid analysis, specific rotation, optical rotatory dispersion of the nickel-complex and the microbiological activity, the product was identical with natural polymyxin B1.View full abstractDownload PDF (645K) -
TOSHIO TAKAGI, TOSHIZO ISEMURA1964 Volume 56 Issue 4 Pages 344-350
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSReduction of all the disulfide bonds in Taka-amylase A by mercaptoethanol in 8M urea yields randomly coiled polypeptide chain. Intact structure of the enzyme can be recovered by removal of the reducing and denaturing reagents and by subsequent airoxidation.
When the reoxidation mixture was free from contamination, the reoxidation and reactivation proceeded very slowly. These changes were found to be markedly accelerated by the addition of some of transition metal ions which have been well known with thiols of small molecular weight to catalyze the oxidation of sulfhydryl groups by molecular oxygen. Among them, copper ion was the most effective. A mechanism of the catalysis was proposed.
Reoxidation of reduced proteins so far reported are presumed to be catalyzed by the contaminating metal ions, since no catalyst has been used. To obtain reproducible results in reoxidation expriments, the amount of the catalyst should be strictly controlled.
The authors thank Sankyo Co. Ltd. for suppling “Taka-diastase Sankyo.” They also wish to thank Dr. K. Hamaguchi and Mr. K. Yutani for helpful suggestions, and Miss A. Kotake for her assistance in ultracentrifugal analysis.View full abstractDownload PDF (1092K) -
VII. Further Purification and Properties of the Gentisic Acid Forming Enzyme SystemMASAYUKI KANDA, HIROSHIGE WATANABE, YUKIYA SAKAMOTO1964 Volume 56 Issue 4 Pages 351-354
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSHomogentisate: O2 oxidoreductase (decarboxylating) and gentisic aldehyde oxidase from a rat liver extract were purified. Homogentisate: O2 oxidoreductase (decarboxylating) may be a flavin enzyme, containing flavin mononucieotide as a cofactor. Gentisic aldehyde is then oxidized to gentisic acid by a general liver aldehyde oxidase.View full abstractDownload PDF (685K) -
IV. Properties of the Oligoribonucleotide PortionJIRO KOYAMA1964 Volume 56 Issue 4 Pages 355-360
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe chemical nature of streptolysin S' was investigated with a purified toxin obtained by zone electrophoresis and DEAE-cellulose column chromatography (4).
1. The oligoribonucleotide portion found in the purified toxin showed the similar ribonucleotide composition to that of oligoribonucleotides used for the toxin formation and confirmed the previous observation that the toxin contained oligoribonucleotides rich inguanylic acid and with a low pyrimidine content.
2. The oligoribonucleotides isolated from the toxin by phenol treatment restimulated the toxin formation.
3. The oligoribonucleotide portion was resistant to various RNA-hydrolyzing enzymes such as RNase T1, RNase T2, RNase I and spleen phosphodiesterase.
4. The DEAE-cellulose column chromatography of streptolysin S' in the presence of urea showed that the oligoribonucleotide portion consisted of heterogeneous components and presented in aggregated state.
5. Although pH 10.5 treatment of streptolysin S' dissociated the aggragated oligoribonucleotide portion, the dissociation did not take place with the original oligoribonucleotides used for the toxin formation.
From the results hitherto obtained, the present author concluded that streptolysin S' was a complex polypeptide containing oligoribonucleotides added for the toxin formation.
The present author wish to express sincere thanks to Prof. F. Egami for his guidance and encouragement during this work. He also express thanks to Mr. Y. Sokawa for his useful discussions.View full abstractDownload PDF (446K) -
III. Amino Acid Composition of Rat Liver CatalaseTOKUHIKO HIGASHI, YASUO SHIBATA1964 Volume 56 Issue 4 Pages 361-363
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe amino acid composition of purified rat liver catalase was determined by ion exchange chromatography using a Beckman Spinco analyzer as follows (in grams of amino acid per 100g. of protein): alanine, 5, 88; glycine, 4.36; valine, 6.76; leucine, 6.93; isoleucine, 4.26; proline, 7.95; phenylalanine, 8.27; tyrosine, 5.67; tryptophan, 2.64; serine, 4.29; threonine, 5.00; cysteine, 0.37; halfcystine, 0.59; methionine, 3.01; arginine, 8.54; histidine, 4.90; lysine, 7.45; aspartic acid, 14.89; glutamic acid, 13.27; amide NH3, 2.05.
The authors wish to thank Prof. N. Shimazono and Associate Prof. H. Hirai of this Department of Biochemistry for their constant encouragements, and are deeply grateful to Mr. S. Horiuchi of the Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo for his performing the amino acid analyses and also to Miss K. Taira of the Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, University of Tokyo for her carrying out the sedimentation analysis.View full abstractDownload PDF (818K) -
YASUO KAGAWA, AKIKO KAGAWA, NORIO SHIMAZONO1964 Volume 56 Issue 4 Pages 364-371
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. The mechanism of the alteration in the enzyme activity was investigated concerning the hexose monophosphate shunt in rat liver. Refeeding of unfed rats on high-carbo-hydrate diet caused the increases of specific activity of the enzymes in the supernatant fraction; G6PDH (+1, 280%), 6PGL (+110%) and 6PGDH (+172%) without increase in LDH. This effect was specific for liver among various tissues. The effect on the dehydrogenases was hardly caused by a high-fat diet and was inhibited by the administration of 8-azaguanine.
2. The increased activities of the three enzymes were stable against protein purification procedures. There was no significant difference in kinetic properties of the enzymes between unfed and refed rats. Participation of activators or inhibitors and the changes in enzyme stability and intracellular distribution in this phenomenon was not detected. Titration of G6PDH with NADPH suggested 30-fold increase of the enzyme in refed livers.
3. It may be concluded that the elevated enzyme activities in this phenomenon are caused by the increase in enzyme protein. The nature of this adaptation was discussed.
The authors express their gratitude to Dr. H. Takiguchi of their laboratory for his assay of 6PGL. Thanks are due to Dr. T. Sugimura for his gift of ethionine and 8-azaguanine and to Dr. M. Ohishi for his gift of puromycin, ribonuclease [EC 2. 7. 7. 16] deoxyribonuclease [EC 3. 1. 4. 5] and amino acids.View full abstractDownload PDF (499K) -
KOSCAK MARUYAMA, YAYOI ISHIKAWA1964 Volume 56 Issue 4 Pages 372-374
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
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MASATAKE HORI, EIKO KONISHI1964 Volume 56 Issue 4 Pages 375-376
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
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TSUYOSHI OHNISHI, HIROÉ KAWAMURA1964 Volume 56 Issue 4 Pages 377-378
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
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MIKI GOTO, TADASHI OKADA, HUGH S. FORREST1964 Volume 56 Issue 4 Pages 379
Published: October 25, 1964
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (90K)
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