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60 巻, 5 号
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NORIKO MURAOKA, SHINJI NAKAJIMA, TAKESHI KATO, TOSHIO SHIMADA1966 年 60 巻 5 号 p. 489-495
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリーA procedure for the crystallization of rabbit liver ferritin was described.
The substance with absorption at 270 mμ, which seems to interfere with the crystallization of ferritin was removed through the purification procedure with acetone.
The spleen was more suitable than the liver for the preparation of rabbit ferritin in crystal form.
Some physicochemical properties of rabbit liver ferritin were described.
The assistance of Miss S. Shinjo, Miss T. Watanabe and Mr. K. Saito is greatfully acknowleged.抄録全体を表示PDF形式でダウンロード (2239K) -
VII. On the Sulfhydryl Groups of the EnzymeEIICHI MISAKA1966 年 60 巻 5 号 p. 496-501
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリーLipoamide dehydrogenase [EC 1. 6. 4. 3] from Saccharomyces oviformis contains 4 moles of free -SH groups per molecule which react with PCMB. The enzyme also contains two catalytic disulfide linkages. Of the 4-SH groups, three react rapidly with PCMB while one reacts slowly. In the presence of 0.5% dodecylsulfate, 6-SH groups per molecule were reacted. This indicates the presence of -SH groups which are not reactive in the absence of the detergent.
When the 3PCMB-sensitive -SH groups were blocked by PCMB, both the lipoamide and the menadione reductase activities were inhibited. After 4-SH groups had reacted with PCMB, however, the menadione reductase activity was restored almost to the level of the native enzyme but the lipoamide reductase activity was not. When the catalytic disulfide as well as the free -SH groups were blocked by PCMB, the lipoamide reductase activity was almost completely inhibited but the menadione reductase activity was slightly increased.
The author wishes to thank Dr. G. Sunagawa, the Director of these Laboratories, for his encouragement and Dr. K. Nakanishi for his valuable advice and discussion.抄録全体を表示PDF形式でダウンロード (483K) -
AKIRA MAKITA, CHIYUKI SUZUKI, ZENSAKU YOSIZAWA1966 年 60 巻 5 号 p. 502-513
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリーGalactosamine-containing glycolipid (Forssman hapten) was isolated from both equine kidney and spleen, by repeated chromatography on silicic acid.
Its chemical composition was identical with that of globoside I (ceramide-glucose-galactose-galactosamine=1:1:2:1 in molar ratio) of human erythrocytes and kidney. The results of methylation of F-hapten followed by gas-chromatographic analyses also indicated a similar structure to that of globoside I.
However, F-hapten could be distinguished from globoside I in its mobility on a thin-layer chromatogram, its optical rotation and its serological specificities.
In serological studies which involved hemolysis- and hemagglutination-inhibition and precipitin reactions, F-hapten isolated from equine kidney and spleen showed human blood group A-activity, in addition to F-activity, while globoside I did not.
The haptens from both equine kidney and spleen were shown to be identical on the bases of their chemical and immunological properties.
The present investigation indicates that the determinant group of F-hapten of the glycosphingoside is most probably O-α-N-acetylgalactosaminoyl-(1→3)-galactose at the non-reducing end, differing from F-inactive globoside I in the anomeric configuration of the disaccharide.
The enhancing effects of various lipids on the F-activity of the hapten were determined by the hemolysis-inhibition test. The results indicated that with an optimal amount of added auxiliary lipid, 0.002-0.018μg. F-hapten/ml. could readily be measured.
The roles of auxiliary lipids and of the lipid moieties in lipid haptens, and the relationship between F-hapten and human blood group-A antigens are discussed.
The authors are indebted to Prof. T. Yamakawa for his kindness in allowing us to continue this research in the present laboratory.
The authors wish to thank Prof. T. Miki and Mrs. Y. Hayashida in Tokyo Medical and Dental University School of Medicine for their guidance in the sheep-red cell hemolysis-inhibition test, for carrying out the human-red cell isoagglutination test, and for gifts of antisera. Thanks are also due to Prof. M. Tomoda of Kyoritsu Pharmaceutical College and Dr. Y. Suzuki of Tohoku University School of Pharma-cology for gifts of authentic samples of 2, 3, 6-tri-O-methylgalactose and egg yolk lecithin, respectively.抄録全体を表示PDF形式でダウンロード (2468K) -
LXXXIII. Metabolism of Deoxycholic Acid and 5β-Cholestane-3α, 12α-diol in the EelSHIGEO HIROFUJI1966 年 60 巻 5 号 p. 514-518
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリーThe metabolism of deoxycholic acid and 5β-cholestane-3α, 12α-diol in the eel was studied, and it was shown that the latter compound was efficiently converted to deoxycholic acid. No evidence for the formation of cholic acid from deoxycholic acid in the eel was obtained.
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 (298K) -
I. Chromatography on a DEAE-Sephadex Column with a Concentration Gradient of Ammonium Sulfate and of Potassium Chloride Containing Organic SolventMASAZUMI MIYAZAKI, MASAHARU KAWATA, SHOSUKE TAKEMURA1966 年 60 巻 5 号 p. 519-525
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリーColumn chromatography of Torulopsis utilis tRNA's with ammonium sulfate-DMFA on DEAE-Sephadex and with organic solvent-KCl systems on DEAE-Sephadex and DEAE-cellulose are described.
The ammonium sulfate-l% DMFA and 10% DMFA-KCl systems on a DEAE-Sepha-dex column were especially useful as an initial step in the large scale preparation of amino acid specific tRNA's. With both systems there was good recovery and several fold purification of the various amino acid-acceptor activities.
The distribution curves of the alanine, valine, glycine and phenylalanine tRNA's suggest that these RNA's are heterogeneous.
The expences of this study were defrayed in part by a grant from the Ministry of Education.抄録全体を表示PDF形式でダウンロード (537K) -
II. Chromatography on a DEAE-Sephadex Column in Phosphate and in BorateMASAZUMI MIYAZAKI, SHOSUKE TAKEMURA1966 年 60 巻 5 号 p. 526-532
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリーChromatography of Torulopsis utilis tRNA on DEAE-Sephadex columns with various concentrations of phosphate or of borate were reported.
Of the solvents tested, M phosphate-5% DMFA and 0.5M borate were the most suit-able for fractionation of tRNA, since they gave good purification and recovery of the various amino acid specific tRNA's. The distribution patterns of the various acceptor activities dif-fered in the M phosphate-DMFA, 0.5M borate, ammonium sulfate-DMFA and DMFA-KGI systems.
Several amino acid-acceptor activities were resolved into multiple peaks especially with the 0.5M borate system.
It was necessary to control the temperature to obtain reproducible elution patterns on chromatography with phosphate or borate, unlike the case with ammonium sulfate.
The expences of this study were defrayed in part by a grant from the Ministry of Education.抄録全体を表示PDF形式でダウンロード (493K) -
II. Substrate Specificity of the EnzymeFUTOSHI IZUMI, KAZUO NAGAI, HIROSHI YOSHIDA1966 年 60 巻 5 号 p. 533-537
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリー1. Hydrolysis of carbamylphosphate and acetylphosphate by an enzyme preparation obtained by treatment of the guinea pig brain microsomes with Nal was remarkably stimulated by addition of K+, and the K+-stimulated hydrolysis was inhibited by ouabain. o-Phosphorylserine, α-glycerophosphate and β-glycerophosphate were not hydrolyzed, and the hydrolysis of ATP was not enhanced by K+.
2. A close similarity was found in the time courses of heat inactivation of K+-dependent phosphatase with carbamylphosphate and p-nitrophenylphosphate as substrates. The reduction of K+-dependent phosphatase activity by heat treatment was lessened in the presence of p-nitrophenylphosphate or carbamylphosphate.
3. The K+-dependent activity of hydro-lyzing carbamylphosphate was far higher than that of hydrolyzing p-nitrophenylphosphate, and the Km values for carbamylphosphate, p-nitrophenylphosphate and acetylphosphate were estimated to be 0.6mM, 1.2mM and 1.6mM, respectively. Accordingly, carbamyl-phosphate was considered to be a better substrate of K+-dependent phosphatase than p-nitrophen ylphosphate.
4. The Km value for K+ with p-nitrophenylphosphate was 2.9mM, whereas the values with carbamylphosphate and acetyl-phosphate were 0.8mM and 1.8mM, respectively. Furthermore it was observed that Li+ was a substitute for K+ when carbamylphos-phate was used as a substrate, though Li+ was less effective with p-nitrophenylphos-phate.
This work was partly supported by a grant from the Japanese Waksman Foundation.抄録全体を表示PDF形式でダウンロード (356K) -
XI. Spectrophotometric Titration of Tyrosyl Groups of Hen and Duck Egg-white LysozymeTAKESHI TOJO, KOZO HAMAGUCHI, MICHIKO IMANISHI, TSUNEHISA AMANO1966 年 60 巻 5 号 p. 538-542
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリーSpectrophotometric titration of the tyrosyl groups of hen and duck egg-white lysozyme [EC 3.2.1.17] was carried out at 245 and 295mμ. It was found that the three tyrosyl groups of hen egg-white lysozyme have pK values of 9.95, 11.6, and 12.6, respectively. In the case of duck egg-white lysozyme, three of the five tyrosyl groups were titrated freely (pK=10.15), and the remaining two ionized with time above pH 12.0 having a pK value of 12.6.
We wish to express our sincere gratitude to Dr. T. Murachi for many helpful discussions.抄録全体を表示PDF形式でダウンロード (358K) -
I. Purification and Some PropertiesHIDEO INOUE, FUJIO SUZUKI, KEIHACHI FUKUNISHI, KOZABURO ADACHI, YOSHIR ...1966 年 60 巻 5 号 p. 543-553
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリーATP citrate lyase [EC 4. 1. 3. 8] was isolated and purified in crystalline form from rat liver, and some of its properties were investigated.
The purified enzyme was homogeneous on sedimentation, on moving boundary electro-phoresis, and on immunological analysis. The molecular weight of the enzyme was estimated to be approximately 500, 000.
The enzyme is highly specific for citrate and ATP, and also requires Mg2+ and sulf-hydryl compounds, such as 2-mercaptoethanol, gultathione or Cleland's reagent, for the reaction. The Km values are 5.8×10-4M for citrate and 1.72×10-4M for ATP at pH 8.4. ADP competitively inhibits the reaction with respect to ATP. The optimal pH is around 8.4.
The purified enzyme is labile on storage. However, the inactivation is effectively prevented by addition of 0.01M 2-mercapto-ethanol together with 0.001M MgCl2, especially under a nitrogen atmosphere. Cleland's reagent is most satisfactory in this respect.
The authors wish to express their thanks to Professor K. Narita of the Institute for Protein Research, for amino acid analysis, and Professor Y. Izumi of the same Institute for sulfur analysis.
Thanks are also due to Professor M. Yoneda and Dr. Y. Fukui, Institute for Microbial Diseases, Osaka University, for immunological procedures. The authors are also indebted to Dr. T. Takagi, Institute for Protein Research of this university, for his aid with electrophoretic analysis.抄録全体を表示PDF形式でダウンロード (1059K) -
TOSHISUKE KAWASAKI, JIRO KOYAMA, IKUO YAMASHINA1966 年 60 巻 5 号 p. 554-560
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリー1. An at-acid glycoprotein (at-AGP) was isolated from rat serum by means of DEAE-cellulose chromatography followed by Pevikon block electrophoresis.
2. This glycoprotein was proved to be homogeneous on polyacrylamide gel electro-phoresis, CM-cellulose chromatography, Se-phadex gel filtration and ultracentrifugal analysis.
3. It contained hexose, methylpentose, amino sugar and sialic acid, the total carbo-hydrate content being as high as 35%.
4. It was shown that α1-AGP from rat and human sera are similar in chemical and physical properties. They also share unusual properties in immunological analysis, starch gel electrophoresis, and Sephadex gel filtration.
The authors are very grateful to Dr. N. Ui, Institute of Endocrinology, Gunma University, for the ultracentrifugal analyses. They are also grateful to Dr. J. E. Scott, Rheumatism Research Unit, Canadian Red Cross Memorial Hospital, England, for revising the manuscript.抄録全体を表示PDF形式でダウンロード (1219K) -
HAJIME SHIGEMATSU, TSURU SHIIO, HIRO-O TAKESHITA, SHIZUE ONODERA, BUNJ ...1966 年 60 巻 5 号 p. 561-567
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリーParticulate fractions prepared from the posterior silkglands of silkworm, Bombyx mori, were used to construct a cell-free incorporation system and to assay the template activity of silkgland RNAs. In the cell-free system with particulate R, pH 8.0- and 8.5-RNAs showed an especially high stimulative effect on the incorporation of glycine into protein. On the other hand, in the cell-free system with particulate M, only insignificant stimulation was caused by these RNA fractions. The nucleotide composition of RNA fractions extracted successively with buffers of different pH values was analyzed. The compositions of pH 7.0-and 7.5-RNAs were found to be similar to that of ribosomal RNA, and pH 8.5- and 9.0-extracts had nucleotide compositions resembl-ing that of the DNA. pH 8.0-RNA possessed a characteristic composition. Up to a certain amount of added RNA, either sRNA or pH 8.0•8.5-RNA stimulated stoichiometrically the incorporation of glycine by the cell-free system with fraction R. The stimulation of glycine incorporation by the pH 8.0-RNA was also observed in E. coli cell-free system. Phenylalanine incorporation, however, was not stimulated by silkgland RNA.抄録全体を表示PDF形式でダウンロード (554K) -
III. Localization of Cytochrome c-552 in the Surface Layer of CellsTAKESHI FUJITA, RYO SATO1966 年 60 巻 5 号 p. 568-577
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリー1. When the cells of E. coli were treated with lysozyme and EDTA in sucrose-Tris-HCl buffer (pH 8.0), spheroplasts were rapidly formed. The spheroplasts thus formed were stable and their cytoplasmic membrane remained almost unimpared in this medium.
2. Cytochrome c-552 was liberated quanti-tatively into the surrounding medium when the cells were converted to spheroplasts. On the other hand, two intracytoplasmic enzymes, i.e., glucose 6-phosphate and glutamate dehydro-genases [EC 1. 1. 1. 49 and 1. 4. 1. 4], remained in the spheroplasts, with little or no activity appering in the medium. From these findings, it was postulated that the cytochrome is located at the cell surface, probably outside the cytoplasmic membrane.
3. Most of cytochrome c-552 in the cells was also released into the medium when the cells were treated by EDTA alone in the sucrose-Tris medium and washed with dilute buffer.
4. The release of cytochrome c-552 on protoplast formation could be observed also in several other species belonging to Enterobacteriaceae.
The authors are indebted to Mr. Y. Imai and Dr. A. Kurono-Yutani for generous gifts of glucose 6-phosphate and lysozyme, respectively.抄録全体を表示PDF形式でダウンロード (742K) -
TAKAO KOBAYASHI, KOHEI OTOMO, KENSUKE SHIMURA1966 年 60 巻 5 号 p. 578-585
発行日: 1966/11/25
公開日: 2008/11/18
ジャーナル フリー1. From electron microscopic observations, it was demonstrated that the ribosome fraction active in amino acid incorporation in a cell-free system from the posterior silkglands of silkworm contained many polyribosomes which had a wide variety in size and shape.
2. The ribosome fraction were further fractionated by sucrose density gradient centrifugation. There were observed two peaks, one lighter and the other heavier. The latter was active in amino acid incorporation in a cell-free system than the former.
3. Observations made by analytical ultracentrifugation have revealed that the heavier and lighter fractions had the sedi-mentation coefficients of about 420S and 76S, respectively.
4. The Protein: RNA ratios of 420S and 76S fractions were 55.0:45.0 and 49.3:50.7, respectively.
5. The possibility that the nascent fibroin might be concerned in the formation of the larger aggregates of ribosomes was discussed.
The authors wish to express their thanks to Dr. Y. Watanabe in School of Medicine, Tohoku University, for his kind allowance to use the analytical ultracentrifuge, and also to Mr. T. Sato in Faculty of Agriculture, Tohoku University, and to Mr. K. Kurita in Naka Factory of Hitachi Ltd. for their technical assistance in electron microscopic observations.
Authors also wish to express their deep appreciation to Dr. T. Kurasawa, chief of the Experimental Sericultural Station of Miyagi Prefecture, for his kind supply of the silkworm.抄録全体を表示PDF形式でダウンロード (2360K) -
MICHITERU YOSHIDA, KOKI YOKOTSUKA, KENSUKE SHIMURA1966 年 60 巻 5 号 p. 586-588
発行日: 1966/11/25
公開日: 2008/11/18
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MITSURU UCHIYAMA, KAZUTAKA MOMOSE, REIKO MOMOSE, SEIICHI OKUI1966 年 60 巻 5 号 p. 589-591
発行日: 1966/11/25
公開日: 2008/11/18
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AKIRA KOTAKI, MINORU HARADA, KUNIO YAGI1966 年 60 巻 5 号 p. 592-594
発行日: 1966/11/25
公開日: 2008/11/18
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KOKI YOKOTSUKA, MICHITERU YOSHIDA, KENSUKE SHIMURA1966 年 60 巻 5 号 p. 595-597
発行日: 1966/11/25
公開日: 2008/11/18
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SHINJI ISHIKAWA1966 年 60 巻 5 号 p. 598-601
発行日: 1966/11/25
公開日: 2008/11/18
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TATSUYUKI KAMIRYO, MASAMICHI TAKAGI, BUNJI MARUO1966 年 60 巻 5 号 p. 602-604
発行日: 1966/11/25
公開日: 2008/11/18
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MAKOTO ENDO, YOSHIAKI NONOMURA, TOMOH MASAKI, IWAO OHTSUKI, SETSURO EB ...1966 年 60 巻 5 号 p. 605-608
発行日: 1966/11/25
公開日: 2008/11/18
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