The Journal of Biochemistry
Online ISSN : 1756-2651
Print ISSN : 0021-924X
52 巻, 2 号
選択された号の論文の12件中1~12を表示しています
  • Changes of Optical Rotation and Susceptibility to Enzymic Proteolysis with Ovalbumin Denatured by Pressure
    CHIEKO SUZUKI, KEIZO SUZUKI
    1962 年 52 巻 2 号 p. 67-71
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
    1. The denaturation of ovalbumin in aqueous solution by pressure up to 10, 000kg./cm.2, at temperatures of 20 and 30°C and at pH between 5.8 and 8.2 was studied with the solubility test, the susceptibility change to, enzymic proteolysis and the optical rotation change.
    2. It was shown that the results obtained by above three methods were consistent within experimental error, and that the pressure denaturation of ovalbumin in aqueous solution started at about 5, 000kg./cm2 and completed at about 9, 000kg./cm2 after being compressed for 5 or 10 minutes.
    3. Optical rotation change by pressure denaturation was considerably smaller than that of heat or urea denaturation.
    4. Although the denaturation of oval-bumin by high pressure is accompanied by the relatively small change of optical rotation, , the denatured protein is fully precipitable in the solubility test and is fully hydrolysable by a proteolytic enzyme. Therefore the configurational change seems to be extensive, but it seems to cause the transformation to β-form.
    5. In view of the negative volume of activation and the negative entropy of activation, it is of no doubt that the binding of water molecules by electrostriction plays an important role.
    The authors wish to acknowledge the interest and encouragement of Prof. W. Jono and thank Drs. J. Osugi and T. Inagami for discussions as well as the generous assistance given by the staff of the Laboratories of Physical Chemistry in Kyoto University and Ritumeikan University. The expense of this study was defrayed in part by a grant from the Ministry of Education.
  • III. Amino- and Carboxyl-Terminal Sequences of Ribonuclease T1
    KENJI TAKAHASHI
    1962 年 52 巻 2 号 p. 72-81
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
    1. The N-terminal amino acid of RNase T1 was quantitatively determined to be alanine both by the DNFB-method and by the PTH-method.
    2. The C-terminal amino acid of the enzyme was quantitatively determined to be threonine both by the hydrazinolysis method and by the carboxypeptidase method.
    The above results indicated that RNase T1 is composed of a single open polypeptide chain.
    3. The N-terminal sequence was investigated both by the stepwise degradation method of E d m a n and by the partial hydrolysis method and the six amino acid sequence from the terminus was deduced as follows; Ala•1/2Cys•Asp (or Asp-NH2)•Tyr•Thr•Asp(or Asp-NH2). The native enzyme as well as the oxidized enzyme was found to be rather resistant to the action of leucine aminopeptidase.
    4. The C-terminal sequence was investigated by the carboxypeptidase method and the following sequence was elucidated: (Ser, Asp, Ala)•Val•Thr. The first two residues were found to be not essential for the enzymatic function of RNase T1, while the following portion was shown to be connected in some manner with the enzymatic function.
    The author wishes to express his sincere thanks to Prof. F. Egami for his guidance and encouragement during this work. He also expresses his gratitude to Sankyo Co. Ltd. for the kind gift of “Takadiastase Sankyo”. The expense of this study was defrayed in part by a grant from the Ministry of Education.
  • I. Toxic Glycolipids of Mycobacterium fortuitum and Atypical Mycobacteria Strain P16
    ICHIRO AZUMA, YUICHI YAMAMURA
    1962 年 52 巻 2 号 p. 82-91
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
    Purified wax was extracted from Mycobacterium fortuitum and atypical mycobacteria strain P16. Highly purified toxic glycolipid was obtained from purified wax by the repeated column chromatographies on sillica gel, silicic acid and magnesium silicate. By means of chemical analysis and toxicity test, it was shown that toxic glycolipids were identical with cord factor of tubercle bacilli. The toxic lipid contents of Mycobacterium fortuitum and atypical mycobacteria strain P16 were lower than those of various tubercle bacilli strains. Acid component obtained from toxic glycolipid of atypical mycobacteria strain P16 was assumed to have the formula C68H136O4.
    The authors express their deep gratitude to Dr. H. Oura, School of Pharmacy, Toyama University, for his helpful criticism and Dr. A. Yasuda and Dr. Y. Fujiwara for cultivation of bacilli. The authors indebted to Mrs. S. Matsuba for elementary analysis, and to Mr. K. Hikita for spectral measurment. A part of this work was supported by a Grant-in-Aid for Scientific Reserch from the Ministry of Education, to which the authors' thanks are due.
  • II. Toxic Glycolipids of Mycobacterium smegmatis, Mycobacterium phlei and Atypical Mycobacteria Strain No. 6 and No. 22
    ICHIRO AZUMA, TOKUKO NAGASUGA, YUICHI YAMAMURA
    1962 年 52 巻 2 号 p. 92-98
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
    1. Purified waxes were extracted from Mycobacterium smegmatis, Mycobacterium phlei and atypical mycobacteria strain No. 6 and No. 22 in the yields of 2 to 4 per cent of bacilli.
    2. Toxic lipids were fractionated from purified wax by column chromatographies on silica gel, silicic acid and Florisil and their yields were 0.3 to 2.2 per cent of purified wax.
    3. These glycolipids were active at 0.01mg. dose levels in the toxicity to mice.
    4. Lipid moiety of these glycolipids were mycolic acids. Mycolic acids of Mycobacterium smegmatis and Mycobacterium phleiii were different from mycolic acid of tubercle bacilli and assumed to have the formula C67H134O6.
    5. Water soluble fraction of these toxic glycolipids was trehalose.
    The authors express their deep gratitude to Dr. H. Oura, School of Pharmacy, Toyama University, for his helpful suggestion and Dr. A. Yasuda and Dr. Y. Fujiwara for cultivation of bacilli. The authors are indebted to Mrs. S. Matsuba for elementary analysis, and to Mr. K. Hikita for spectral measurement.
  • KEI NAGANO, MAKOTO NAKAO
    1962 年 52 巻 2 号 p. 99-102
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
    1. Regaining of potassium by cold-preincubated red cells was suppressed by 3×10-8 to 3×10-6M ouabain, while turnover of the phosphate moiety of phospholipids of red cell membrane, especially of phosphatidic acid, was not inhibited by ouabain.
    2. Erythrocyte ATPase was inhibited by ouabain in the same range of concentration.
    3. ATP added to the plasma inhibited potassium resorption from the plasma into the red cells.
    4. Based on these results, a possibility was discussed for ATPase as playing a role in cation transport of red cells.
  • III. Further Purification and the Homogeneity of a Fragment with Peptic Activity
    KIYOCHIKA TOKUYASU, MASARU FUNATSU
    1962 年 52 巻 2 号 p. 103-107
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
    1. Further purification of the partially purified fragment of pepsin was attempted using a column of DEAE-cellulose and/or CM-cellulose and resulted in the separation of two components, a major enzymatically active component-Fragment A-and a minor inactive component.
    2. Fragment A thus obtained behaved as a homogeneous protein ultracentrifugally, chromatographically, and electrophoretically.
    3. Approximately one third of the total peptic activity produced in the dialysate by dialysis autolysis of pepsin (0.4 per cent of that of starting material) obtained in Frag-ment A and 1.7 per cent of total nitrogen produced in the dialysate (0.6 per cent of that of starting material) was obtained in Fragment A.
    4. The specific activity of Fragment A against casein or bovine hemoglobin corresponded to approximately half of that of crys-talline pepsin.
    We wish to express our thanks to Dr. Martin Sonenberg, Head, Metabolism Laboratory, Sloan-Kettering Institute for Cancer Research, for his help during preparation of the manuscript.
  • II. Metabolism of Glucose
    ISAMU SHIIO, SHIN-ICHIRO ÔTSUKA, NOBORU KATSUYA
    1962 年 52 巻 2 号 p. 108-116
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
    1. Glucose was completely oxidized to form 6 moles of carbon dioxide in the washed cells of Brevibacterium flavum No. 2247, grown in a biotin-rich medium, while, in the cells grown in a biotin-poor medium, 0.5-0.7 moles of α-ketoglutarate as well as carbon dioxide were formed as the end product. The end products in the aerobic metabolism of pyruvate were similar as in that of glucose, respectively.
    2. Any significant effect of biotin concentration in the culture medium was not observed on the formation of pyruvate and citrate by the washed cells, on the metabolism of citrate in the detergent-treated cells, and also on the various enzyme activities of cell extracts.
    3. A considerable amount of L-glutamate was formed from glucose also in the cells grown in a biotin-rich medium if some detergents were added to the reaction mixture; the detergents had been known to make these cells permeable to glutamate.
    4. These results support strongly the “permeability hypothesis” that has been previously proposed with regard to the effect of biotin on the extracellular formation of glutamate.
    The present authors are indebted to Dr. H. Oeda and Dr. Y. Tsuchiya of our laboratory for their interest and encouragement during the course of this work.
  • IV. Cytochrome b2 and Yeast L-Lactic Dehydrogenase
    JINPEI YAMASHITA, KAZUO OKUNUKI
    1962 年 52 巻 2 号 p. 117-124
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
    1. Sedimention constant, diffusion coefficient and iron content of cytochrome b2 were measured. The sedimentation constant was determined to be 2.2 S, the diffusion coefficient 10.8×10-7cm2. sec.-1, and iron content 0.24 per cent. The molecular weight of cytochrome b2 was calculated as 18, 000 from the physicochemical values, and as 22, 000 from the iron content.
    2. Cytochrome b2 had absorption maxima at 275, 359, 411 and 534mμ in its oxidized state and at 422, 528 and 557mμ in its reduced state, the molar extinction coefficient at 557 my was found to be 37.9×103M-1cm.-1. The absorption maxima of the reduced pyridine haemochromogen of cytochrome b2 were at 419, 528 and 557mμ.
    3. In neutral solution CO had no effect on the absorption spectrum of reduced cytochrome b2, but in acidic solution, it combined with CO, giving absorption maxima at 419, 543 and 575mμ. Cytochrome b2 appeared to have no affinity for CN- or N3-.
    4. In acidic and alkaline solution, the methylene blue- and cytochrome c-reducing activity of yeast L-lactic dehydrogenase were lost more rapidly than the phenazine methosulphate-reducing activity.
    From these findings, a possibility was discussed that yeast L-lactic dehydrogenase consists of a dehydrogenase moiety and a haem protein moiety.
    The authors would like to express their thanks to Prof. B. Hagihara of this university for technical help in the use of the oxygen electrode and to Mr. K. Fujii (Oriental Yeast Co., Ltd., Osaka) for kindly supplying large quantities of baker's yeast.
  • XLVI. Isolation of a New Bile Sterol, 3α, 7α, 12α-Trihydroxy-26, 27-Epoxycholestane from Carp Bile
    TAKAHIKO HOSHITA
    1962 年 52 巻 2 号 p. 125-130
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
    1. Four kinds of bile sterols were isolated from carp bile by use of reversed phase partition chromatography.
    2. It was verified that chemical constitution of one of the new bile sterols corresponds to 3α, 7α, 12α-trihydroxy-26, 27-epoxy-cholestane.
    The author wishes to express his deep gratitude to Prof. T. Kazuno for his kind advice throughout this work. He also thanks Prof. G. A. D. Haslewood of Guy's Hospital Medical School, London for his generous help in identification of allocholic acid.
  • EIJI ITAGAKI, TAKESHI FUJITA, RYO SATO
    1962 年 52 巻 2 号 p. 131-141
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
    A preparation containing formate dehydrogenase, cytochrome b1 and nitrate reductas_??_ was solubilized and partially purified from the particulate fraction of E. coli cells growr_??_ anaerobically under the conditions favorable for nitrate respiration. Some properties o_??_ formate dehydrogenase and cytochrome b1 in this purified preparation were investigated It was suggested that formate dehydrogenase is a metalloflavoprotein with essential sulf_??_ hydryl groups. Its activity was strongly bu_??_ reversibly inhibited by molecular oxyge_??_ The prosthetic group of cytochrome b1 was decisively identified as protoheme, and it was found that the cytochrome is autoxidizable. The reduction of cytochrome b1 by formate was shown to require, in addition to formate dehydrogenase, a lipid-soluble factor which could be replaced by vitamin K3 in the solubilized system.
    We are indefted to Drs. H. Maeno and S. Sakakibara for generous gifts of crude snake venonand crystalline protohemin, respectively.
  • ICHIRO AZUMA, YUICHI YAMAMURA
    1962 年 52 巻 2 号 p. 142-144
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
  • TSUYOSHI OHNISHI
    1962 年 52 巻 2 号 p. 145-147
    発行日: 1962/08/25
    公開日: 2008/11/18
    ジャーナル フリー
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