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50 巻, 5 号
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AKIO MAEDA1961 年 50 巻 5 号 p. 377-385
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリーThe ribosomal RNA prepared from yeast shows two distinct peaks in the sedimentation pattern. The relative ratio of the amount of those two components changes as the function of the amount of salt, especially of magnesium ions. In the presence of 0.1M of NaCl, some portion of the fast sedimenting component converts to slower one as the consequence of removal of magnesium ions. The former is almost completely converted to the latter by further removal of salt by dialysis against distilled water. In this state, the RNA shows very high viscosity, low dextro-rotation and high degree of hyperchromicity.
Considerable amount of magnesium ions binds to the RNA so that they are not dialyzable against distilled water. The amount of magnesium atom equivalents in such form is about 0.4 per mole of phosphate of the RNA. Below this critical amount, a marked hyperchromicity is ovserved in water. Even in the presence of critical amount of magnesium ions, some increase of the slower peak and increase of viscosity are observed without addition of any other salt.
From the above mentioned results, or-ganization of the RNA in yeast ribosomal particle is discussed.
The auther wishes to express his thanks to Prof. F, Egami of Tokyo University for his interest and, encouragement throughout this work, The author is much indebted to Dr. S. Osawa of the Biological Institute of Nagoya University for his critical reading of manuscript and kind advice in the course of this work. Thanks are also due to members of the staff of the Laboratory of Polymer Science, Physical In-stitute of Nagoya University for their valuable advice and discussion.
The work was supported by the Scientific Re-search Fund of the Ministry of Education.抄録全体を表示PDF形式でダウンロード (2506K) -
TERUHIKO BEPPU1961 年 50 巻 5 号 p. 386-388
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリーA diphosphopyridine nucleotide-linked α-ketoglutarate reductase has been found from Achromobacter sp. The enzyme catalyzes reversible conversion between α-ketoglutarate and L-a-hydroxyglutarate, but the equilibrium of the reaction is very restricted to the direction of formation of the latter. The reaction proceeds at maximal velocity between pH 6-8 in phosphate buffer. Half maximal rate was obtained at a DPNH concentration of 1.3×10-4M and a α-ketoglutarate concentration of 5×10-5M. The enzyme seemed to be specific for DPN and L-α-hydroxyglutarate. The enzyme was inhibited by p-chloromer-curibenzoate.
The author wishes to express his gratitude to Prof. K. Arima, for his kind advice during the course of this work.抄録全体を表示PDF形式でダウンロード (216K) -
III. Reconstruction of Laccase from Its Protein and CopperTSUNEO OMURA1961 年 50 巻 5 号 p. 389-393
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリー1. The copper of laccase was removed from the enzyme by dialysis against acid and cyanide, and colorless apo-enzymes were obtained.
2. The catalytic activity of the apo-enzyme prepared by acid dialysis was not restored by the addition of cupric and cup-rous ions.
3. The catalytic activity of cyanidedialyzed apo-laccase was partly restored by the contact with cuprous ion, but cupric ion was ineffective. The characteristic blue color of laccase was also partly restored by the recombination of apo-enzyme and cuprous ion.
The author wishes to express his hearty gratitude to the late Prof. A. Nishimura for his interest and encouragement during this investigation, and to Prof. Y. Ogura, University of Tokyo, for his valuable advices. Thanks are also due to Mr. S. Yamamoto, the Institute for Infectious Diseases, University of Tokyo, for sedimentation measurements, and to Miss A. Kurono for her co-operation in this work. The author also expresses his gratitude to Saito Co. Ltd. for the kind supply of lacquer latex.抄録全体を表示PDF形式でダウンロード (1261K) -
IV. Purification and Properties of a Blue Protein Obtained from Latex of Rhus verniciferaTSUNEO OMURA1961 年 50 巻 5 号 p. 394-399
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリー1. A new blue protein was isolated from the latex of Japanese laquer tree (Rhus vernicifera), and purified by column chromatography on an ion-exchange resin. The purified preparation was electrophoretically and ultracentrifugically homogeneous.
2. The blue protein was found to contain 1 atom of firmly bound copper per molecule. The absorption spectrum of the oxidized from has three absorption maxima (850, 608, and 450mμ) in the visible and near infra-red region, and a maximum (280mμ) in the ultraviolet region. The three absorption peaks in the visible and near infra-red region disappeared on reduction.
3. The blue protein could be reversibly reduced and oxidized by L-ascorbic acid and potassium ferricyanide respectively. The autoxidation of the reduced form by atmospheric oxygen was, however, very slow indicating the lack of oxidase activity.
The author wishes to express his hearty gratitude to the late Prof. A. Nishimura for his interest and encouragement during this investigation, and to Prof. Y. Ogura, U niversity of Tokyo, for his valuable advices. Thanks are also due to Mr. S. Yamamoto, the Institute for Infectious Diseases, University of Tokyo, for sedimentation measurements, to Miss K. Watatsu the Institute of Applied Microbiology, University of Tokyo, for the operation of Spinco model H electrophoresis apparatus, and to Miss K. Ichino for her co-operation in this work. The author also expresses his gratitude to Saito Co. Ltd. for the kind supply of lacquer latexes.抄録全体を表示PDF形式でダウンロード (1269K) -
TSUNEO OMURA, TAKESHI FUJITA, FUMIYUKI YAMADA, SUSUMU YAMAMOTO1961 年 50 巻 5 号 p. 400-404
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリー1. An isoelectric precipitation procedure for the purification of hemocyanin from the blood of Ommatostrephes sloani pacificus was described.
2. Purified hemocyanin obtained by the procedure was electrophoretically and ultracentrifugically homogenous at pH's between 7.0 and 8.5 in 0.1M phosphate buffer. In more acidic media, an associated component was reversibly formed in an equilibrium with the dissociated form.
3. The molecular weight of the dissociated component was 6.13×105.
4. Purified hemocyanin contained 0.260% copper, and traces of silver were found to be always present.
The authors wish to express their hearty gratitude to the late Prof. A. Nishimura for his interest and encouragement during this investigation, and to Prof. Y. Ogura, University of Tokyo, for his valuable advices. Thanks are also due to Miss K. Watatsu, the institute of Applied Microbiology, University of Tokyo, for the operation of Spinco model H electrophoresis apparatus. The authors also express their rgratitude to Messrs. K, Mori, H. Hiyoshi, and M. Kawaguchi, the Fisheries Experimental Station of Shizuoka-ken, for their kind aid in obtaining fresh-caught, living squids.抄録全体を表示PDF形式でダウンロード (1104K) -
I. The Inhibition of Cyclic Phosphodiesterase Activity of Bovine Pancreatic Ribonuclease by Several Substrate AnaloguesTYUNOSIN UKITA, KEIZO WAKU, MASACHIKA IRIE, OTOMATSU HOSHINO1961 年 50 巻 5 号 p. 405-415
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリーThis study was undertaken to under-stand better the mechanism underlining the inhibitory action of nucleotides on ribo-nuclease-IA activities. A number of 2(3H)-pyrimidinones, purine- and pyrimidine-nucleo-sides as well as corresponding nucleotides were tested as to their possible inhibition of RNase-IA activity using 2': 3'-cyclic cytidylate or uridylate as a substrate.
The representative compounds of each group showed a competitive type of inhibition to the enzymatic reaction. The 2(3H)-pyrimidinones, possessing a structure similar to the basic residue of the substrate nucleotides, exhibited a strong inhibitory activity. This was largely increased by substitution of pentafuranosyl residue at the N3-position of these pyrimidinones. The pentafuranosyl structure was the most suitable glycosyl residue in the nucleoside for the inhibition of RNase-IA, however no inhibitory activity was observed for all of the purine ribofuranosyl nucleosides tested.
The phosphoryl group substitution at the sugar residue of nucleosides was found signi-ficant in the enhancement of the inhibitory function of the parent compound, however, this group did not appear to play an important role in the specific affinity of the parent nucleoside to the enzyme. The location of the phosphoryl group in the sugar residue, however, appeared important for enhancement of the inhibitory activity, and the greatest affinity of the nucleotide to the enzyme was realized when the phosphoryl group was attached at the 2'(or 3') hydroxyl group of the pentafuranosyl group.
These observations reveal that in the inhibition of cyclic phosphodiesterase activity of RNase-IA by a nucleotide, the structure of each of its three partial residue (the basic group, the sugar residue and the position of phosphoryl substitution) is an important factor in its affinity to an active site of the enzyme molecule. The basic group seems to have the greatest specificity of the three, while the phosphoryl group is somewhat nonspecific in its affinity to the enzyme.
The authors are indebted to Prof. T. Yamada and D. Mizuno of this Faculty and Prof. Y. Mizuno of the University of Hokkaido for their gifts of test compounds.抄録全体を表示PDF形式でダウンロード (797K) -
HARUHIKO ANDO, ICHIRO FUWA1961 年 50 巻 5 号 p. 416-418
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリー1. The effects of cyanamide on alcohol dehydrogenase and aldehyde dehydrogenase were studied.
2. Cyanamide did not affect the activity of alcohol dehydrogenase.
3. Cyanamide inhibited aldehyde dehydrogenase noncompetitively to acetaldehyde as well as to the coenzyme.
The authors wish to thank Dr. K. Yagi for his guidance and valuable criticisms.抄録全体を表示PDF形式でダウンロード (174K) -
MINORU KAWADA, KAZUO YAMADA1961 年 50 巻 5 号 p. 419-423
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリー1. Lactonase II located in the microsomes was studied. Its optimum pH was at about 7.2, and it was proved to be a SH enzyme.
2. Lactonase II was shown to be very labile against heat and alkali treatment and aging.
3. Lactonase II seemed to act only on D-glucurono-γ-lactone, but not on D-man-nurono-γ-lactone and L-idurono-γ-lactone. It seemed not to act on aldonolactone.
4. The significance of this enzyme in the L-ascorbic acid biosynthesis was discussed.
The authors wish to express their gratitude to Prof. N. Shimazono and Dr. Y. Mano for their helpful advices and continuous encouragement.抄録全体を表示PDF形式でダウンロード (354K) -
SHIRO AKABORI, KOZO NARITA, YASUO FUJII, JUNKO ISHII1961 年 50 巻 5 号 p. 424-427
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリーIt was shown that 0.5% dimethylalloxan in ethanol was useful as a detecting reagent for paper chromatography of amino acids and peptides. On heating the paper chromatogram which was sprayed with the dimethylalloxan reagent, amino acids appeared as stable pinkish red spots. Sensitivity of the reagent was almost the same or slightly less than that of the ninhydrin reagent for most amino acids occurring in proteins. Stability of the colored material formed on a paper chromatogram was far better, than that with ninhydrin. However, the pinkish red color was quite unstable in aqueous solution and no suitable conditions for the quantitative estimation of amino acids in solution could be found.
The present work has been aided in part by Tanabe Essential Amino Acids Research Foundation for which the authors are grateful.抄録全体を表示PDF形式でダウンロード (320K) -
SADAMU NAGAI1961 年 50 巻 5 号 p. 428-433
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリーThe enzymes catalyzing the hydrolysis of N-palmitoyl-DL-valine, N-palmitoyl-L-phenyl-alanine, and N-palmitoyl-L-aspartic acid, besides N-acetyl-DL-valine, N-chloroacetyl-L-phenylalanine, and N-acetyl-L-aspartic acid, have been extracted and partially purified from Mycobacterium avium.
In studies on the properties of the hy-drolyzing activities, it was evidenced that the enzymes for N-palmitoyl-amino acids were distinguished from the enzymes for N-acetyl-or N-chloroacetvl-amino acids.
The author wishes to express his sincere gratitude to Prof. Akabori of Osaka Univ. and Prof. Yamamura of Kyushu Univ, for their constant interest and encouragement.抄録全体を表示PDF形式でダウンロード (399K) -
IV. Fractionation of Habu Snake Venom by Chromatograpy on CM-CelluloseHIROO MAENO, SUSUMU MITSUHASHI1961 年 50 巻 5 号 p. 434-439
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリーThe column chromatography of a venom of Japanese venomous snake, Habu (Trimeresurus fiavoviridis), on carboxymethyl cellulose revealed that at least five diesterases containing two RNases and one DNase are present, all of which were separable from lethal toxicity.
Phosphomonoesterase and proteolytic enzymes may also be separable from the toxicity.
There were found at least three 5'-nucleotidase fractions which were chromatographically separable from each other. Two of these nucleotidase fractions run parallel to lethal toxicity.
The authors are indebted to Prof. F. Egami, Department of Biophysics and Biochemistry, the University of Tokyo, for his helpful advice.抄録全体を表示PDF形式でダウンロード (442K) -
I. Purification and Properties of Cytochromes b4 (I) and b4 (II)KATSUJI HORI1961 年 50 巻 5 号 p. 440-449
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリー1. Five chromoproteins: cytochrome c551, brown protein, cytochromes b4 (I), b4 (II) and HdR (cyt. 625, 553) were obtained in soluble form from a halotolerant Micrococcus and were purified by chromatography on DEAE-cellulose column.
2. Reduced cytochrome b4 (I) exhibited an absorption spectrum with double α bands at 554 and 548mμ while reduced cytochrome b4 (II) had single a band at 554-555mμ.
3. Cytochrome b4 (I) had two heme groups per molecule, on the other hand, cyto-chrome b4 (II) had one heme group per molecule.
4. Oxidation-reduction potential for cyto-chromes b4 (I) and b4 (II) were +0.113 volt. and 0.180 volt., respectively.
5. These cytochromes exhibited c-type pyridine hemochromogen spectra and their heme groups did not split from the proteins by treatment with HCl-acetone and also HCl-methylethylketone.
6. They were reduced by DPNH, succinate and formate and were rapidly reoxidized by the addition of nitrate, nitrite and hydroxylamine in the presence of crude extracts of the Micrococcus. Oxygen could also reoxidize them but at a slower rate.
7. A possible scheme of electron transfer from DPNH, succinate and formate via cyto-chrome of type-b, cytochromes b4 (I), b4 (II) and c551 to nitrate, nitrate, hydroxylamine and oxygen was presented.
The anther wishes to express his gratitude to Prof. F. Egami and Dr. S. Taniguchi for their valuable and encouraging advices and to Mr. E. Itagaki for his generous discussions throughout this work. The auther also expresses his thanks to Prof. K. Okunuki and Dr. I. Sekuzu for their useful suggestions during the course of the purification of cytochromes. The auther is indebted to Prof. N. Ui for the measurements of the molecular weights of cytochromes, to Prof. T. Mori for the spectroscopic observations of cytochromes and to Nagoya Factory of Fujisawa Chemicals Co. Ltd, for the cultivation of aerobic and anaerobic cells of the bacterium.
This research was supported in part by a grant from the Scientific Research Fund of the Japanese Ministry of Education.抄録全体を表示PDF形式でダウンロード (619K) -
YOSHIAKI NOSOH1961 年 50 巻 5 号 p. 450-457
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリーA new azomercurial, sodium 4-(4-acetoxy-mercuriphenylzao)-1, 7-Cleve's acid (abbreviated to MPAC) was synthesized, and its applicability for the spectrophotometric and quantitative determination of sulfhydryl groups of mercaptans and proteins was studied under various experimental conditions. The dye, when used with glycine in solution, has a high specificity of reacting only with sulfhydryl groups, and its application to mercaptans and proteins gave us the same contents of sulfhydryl groups as observed by other techniques. The highly specific reaction of MPAC in the glycine-acetate buffer was due to the formation of a MPAC-glycine complex, in which the glycine residue having an appropriate affinity with the mercury atom suppressed the non-specific reactions with ions or radicals having a weaker affinity with the mercury atom. The dye in its application has two major advantages; i) we observe the spectral change in the visible region, where most proteins and extracts from biological materials absorb light weakly and ii) the solubility of the dye in water or buffers is much higher than those of the mercurials so far synthesized, so that we may obtain a greater reading of absorbance change brought about by mercaptide formation.
The author wishes to express his gratitude to Prof. K. Shibata for his valuable advices and encouragement throughout this investigation.抄録全体を表示PDF形式でダウンロード (551K) -
I. Transfer of Radioactivity from Prelabelled Cell Debris to Particulate Fractions in the Cell Free Systems of the Posterior SilkglandYOSHIAKI MIURA, HIROWO ITO, SHIGEO TANAKA, KENHIKO MOMOSE, KIYOSHI SUN ...1961 年 50 巻 5 号 p. 458-466
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリーThe investigation of the transfer of radio-activity from prelabelled cell debris to particulate fractions in the cell-free prepara-tions of the posterior silkgland was developped.
1. After labelling of silkglands with C14-glycine for 15 minutes in vivo or in vitro, posterior silkglands were collected and homogenized. The cell debris fraction was obtained by centrifugation at 700×g for 10 minutes. Microscopic observation of cell debris revealed that major components were nuclear debris and fragments of cell membranes without any intact cells.
2. Prelabelled cell debris thus prepared was highly radioactive and it was shown that under appropriate conditions, the radioactivity was transferred enzymatically to particulate protein. This was confirmed by dinitrophenylation study of isolated particles after the reaction.
3. Radioactivity was principally incorporated into protein of particulate fractions, but not very much into lipid and supernatant fractions.
4. As regards the transfer reaction, amino acid activating enzyme (E3) seemed to be unnecessary, whereas amino acids mixture, GTP, ATP and ATP-generating systems were required.
The work has been supported by grants partly from the Ministry of Education to the research group for Fibroin Biosynthesis and partly from the Rockefeller Foundation to one of the authors. We wish to thank Dr. K. Shimur a and his staff for their valuable discussion and Dr. B. Maruo for his kind supply of C14-all-labelled amino acids.抄録全体を表示PDF形式でダウンロード (1264K) -
KIMIKO ASANO, FUJIO EGAMI1961 年 50 巻 5 号 p. 467-470
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリー1. Yeast s-RNA was slowly, but finally as well digested as commercial yeast RNA by RNase T1
2. Amino acid-charged oligonucleotide was obtained by the digestion of C14-labeled amino acid-charged s-RNA by RNase T1.
3. The existence of an amino acid ter-minal nucleotide sequence (-GpApCpCpA-amino acid) was suggested.
The authors are indebted to Dr. S. Osaw a of Faculty of Science of Nagoya University for his helpful advice and to Sankyo Pharmaceutical Co. Ltd. for the gift of Takadiastase Sankyo.抄録全体を表示PDF形式でダウンロード (255K) -
TAKASHI HASHIMOTO, YOSHINARI ISHII, MASAMITI TATIBANA, HARUHISA YOSHIK ...1961 年 50 巻 5 号 p. 471-472
発行日: 1961/11/25
公開日: 2008/11/18
ジャーナル フリーPDF形式でダウンロード (144K)
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