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Volume 73 (1973)
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- Issue 2 Pages 199-
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Volume 73, Issue 2
Displaying 1-27 of 27 articles from this issue
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IV. Activity and FAD-fluorescence Change Caused by the Chemical Modification of Tryptophyl and Tyrosyl Residues in Glucose OxidaseHaruhito TSUGE, Hisateru MITSUDA1973 Volume 73 Issue 2 Pages 199-206
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESS1. Apoenzyme of, glucose oxidase [β-D-glucose: O2 oxidoreductase, EC 1. 1. 3. 4] prepared with the usual acid-ammonium sulfate method was rechromatographed on a Sephadex G-200 column equilibrated with 0.1 M sodium phosphate buffer (pH 7.0) containing 1 mm dithiothreitol, and an electrophoretically homogenous preparation was obtained.
2. Incorporation of FAD into apoenzyme of glucose oxidase which had been modified with 2-hydroxy-5-nitrobenzyl bromide was inhibited during the first 20 min in the reconstitution reaction.
3. N-Bromosuccinimide titration of holoenzyme for tryptophan at pH 5.6 and 4.0, in the presence and absence of 4.8 M urea, resulted in the increase in FAD fluorescence, while N-acetylimidazole titration for tyrosyl residue only slightly increased the FAD fluorescence.
4. Evidence for the interaction between the isoalloxazine moiety of FAD and the apoenzyme was obtained from fluorometric studies on the chemical modification of apo- and holo-enzyme.View full abstractDownload PDF (4169K) -
Koko MURAKAMI, Sadahiko ISHIBASHI1973 Volume 73 Issue 2 Pages 207-215
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSHexokinase [EC 2.7.1.1] isoenzyme pattern of Ehrlich ascites-tumor cells was found to be variable. Sometimes, Types I and II were detected in almost equal activity, while in other cases Type I existed almost solely. Incubation of the cells in vitro in Krebs' saline serum substitute caused an increase in the proportion of Type II irrespective of the original isoenzyme pattern, while such an increase was hardly observed when glucose was omitted from the medium. Hexokinase activity measured by the usual method tended to increase in the former case, whereas it decreased in the latter. It was assumed that the hexokinase isoenzyme profile was dependent on the availability of glucose, but glucose concentration in the ascites fluid in situ was almost nil in all cases. Intraperitoneal injection of 2-deoxyglucose caused marked increase in the glucose level in the ascites fluid as well as in the proportion of Type II in the cells. The increase in the latter, however, would not be simply attributable to the increase in the former, since incubation of the cells in the medium containing 2-deoxyglucose instead of glucose induced more increase in Type II as compared with the cases of the incubation in the usual medium containing glucose. It was assumed from these facts that glucose metabolism was greatly involved in the regulatory mechanism for hexokinase Type II in Ehrlich ascites-tumor cells. However, the regulation may be done not by glucose itself but by some other factor (s) being concerned with or produced in the glucose metabolism. Possible mechanism was discussed for the regulation.View full abstractDownload PDF (12983K) -
Tadashi ASAHI, Mikio NISHIMURA1973 Volume 73 Issue 2 Pages 217-225
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSThe mitochondrial and cytoplasmic forms of malate dehydrogenase [EC 1.1. 1.37] were purified from the cotyledons of mung bean. There was no significant difference in pH optimum between the isoenzymes.Double reciprocal plots were linear for all substrates and coenzymes except for oxalacetate at extremely high concentrations. Some differences in the Michaelis constants were observed between the isoenzymes. However, the differences did not seem very important from physiological viewpoints, and moreimportant differences were observed in the regulatory properties. NADH was a competitive inhibitorfor NAD+-reducing activity of the cytoplasmic enzyme with regard to NAD+and L-malate. For the activity of the mitochondrial enzyme, NADH was a competitive inhibitor with regard to NAD+ but changed the saturation curve of L-malate from a hyperbolic to a sigmoidal curve.ATP was a strong inhibitor for NAD+-reducing activity of the mitochondrial enzyme but notfor the activity of the cytoplasmic enzyme.View full abstractDownload PDF (1177K) -
II. Comparison of Activity of Mitochondrial and Cytoplasmic L-Glycerol-3-phosphate Dehydrogenases in Testes and Liver of Developing Male Japanese QuailMasahiro YAMADA1973 Volume 73 Issue 2 Pages 227-233
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSThe activities of mitochondrial and cytoplasmic L-glycerol-3-phosphate dehydrogenases [EC 1. 1. 99. 5 and EC 1. 1. 1. 8] from testes and liver were compared in developing male quail, Coturnix coturnix japonica.
The Km values for L-glycerol 3-phosphate of the mitochondrial enzyme from testes and liver were determined as 2.5 and 12.5 mM, respectively, at pH 7.5 and 30°C.
The highest specific activities of the cytoplasmic enzyme and the mitochondrial enzyme of liver were obtained about 12 and 21 days after hatch, respectively, in the developing stage.
The highest specific activity of the mitochondrial enzyme of testes was obtained 56 to 64 days after hatch, while that of the cytoplasmic enzyme was obtained in a grown up adult male.
Ratio of specific activity of the mitochondrial enzyme to that of the cytoplasmic enzyme of testes was remarkably different from that of liver during the developmental stage. The ratio in testes of adult male was about 100-fold than in liver.View full abstractDownload PDF (1053K) -
Masao ONO, Hiroshi MARUTA, Den'ichi MIZUNO1973 Volume 73 Issue 2 Pages 235-243
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSConcanavalin A (Con A), a plant hemagglutinin, was converted to an insoluble derivative by coupling with Sepharose particles using the cyanogen bromide method. Induction of DNA synthesis in vitro by this Sepharose-Con A complex was measured in terms of incorporation of tritiated thymidine into acid insoluble materials in rabbit spleen cells.
The results were as follows. The Sepharose-Con A complex showed mitogenic activity, but its activity was approximately 14% of that of free Con A. The rates of incorporation of tritiated thymidine on addition of the complex and of free Con A were the same, but the complex resulted in higher maximum incorporation. a-αMethyl D-mannoside (α-MM) had no -protective effect on the coupling reaction of the Sepharose-Con A complex at molar ratios of α-MM to Con A of up to 104. The interaction between the Sepharose-Con A complex and spleen cells did not cause agglutination.
It is concluded that the initial information of mitogenic activity is transmitted to rabbit spleen cells without the entrance of Con A into the cells, and consequently, that Con A interacts first with the cell surface membrane.View full abstractDownload PDF (3871K) -
Isao KATSURA, Haruhiko NODA1973 Volume 73 Issue 2 Pages 245-256
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSSupplementing our previous report (J. Biochem., 69, 219 (1971)), the formation of reconstituted myosin filaments was studied.
1) The concentration of monomers in equilibrium with the filaments was independent of the length of filaments and of pH at which filaments were made. The changes inthe monomer concentration by changing pH and KC1 concentration were measured in the range of pH 6-8and 0.195-0.24 M KC1.
2) The length of filaments made by fast dilution was independent of protein concentration down to a very low concentration of 5 μg/ml.
3) The length of filaments made by dialysis was longest at pH 7 and decreased on both sides.
4) RNase [EC 2. 7. 7.16] treatment and partial removal of the light chains of myosin did not change the length of filaments, whereas LiC1treatment and repeated precipitation shortened it. Removal of an unknown factor from myosin by DEAE-SephadexA-50 shortened the length of filaments in the case of slow dilution and addition of the factorto the purified myosin elongated filaments to the original length. Some properties of the factorwas studied.View full abstractDownload PDF (7018K) -
Isao KATSURA, Haruhiko NODA1973 Volume 73 Issue 2 Pages 257-268
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSElectron microscopic studies of light meromyosin aggregates were performed in a solution of potassium chloride covering a wide range of pH and ionic strength. In addition to various types of aggregates already reported, some new types of aggregates were found. The fibers of 430 Å period adhered with each other with a stagger of 3/8 period, thus indicating that the interaction between filaments was different from the interaction between molecules within a filament. Based on the observation of the negative staining pattern of 430 Å fibers and from that of a thin carrot-shaped, fiber, a “tile-roof” model of the fiber was proposed. The thickness of fibers measured by shadow casting was almost the same as their width, while the thickness of sheets was 70-140 Å and independent of their width (0.1-1 μm). The thickness of square nets was usually about 70 Å at the periphery but much thicker at the central part. Both the strands of square nets (70 Å inwidth) and a new type of fiber consisted of fibrils of about 35 Å in width. Fibers were formedat pH lower than 7.5 while both sheets and square nets were formed at pH higher than 7.0. The factorwhich determined the kind of formation whether sheets or square nets was the aging of myosin solution from which light meromyosin was prepared and the duration of digestion.View full abstractDownload PDF (20929K) -
Tomoko FUJII, Masaru KAWAMURA, Keiichi YAMAMOTO, Koscak MARUYAMA1973 Volume 73 Issue 2 Pages 269-280
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSInteraction of troponin I, one of the components of the calcium receptive protein complex of muscle, troponin, and F-actin was investigated with a number of physicochemical techniques. At concentrations of KC1 between 0.03 and 0.06 M and at pH 6.5-8.5, F-actin formed a complex with troponin I and produced an easily precipitable aggregate. Flow birefringence studies suggested that troponin I formed a large nonbirefringent aggregate of F-actin. In the presence of a small amount of troponin I (up to20% of F-actin by weight), F-actin could be dispersed into shorter particles after sonication. The Mg-activated ATPase [EC 3. 6. 1. 3] activity of actomyosin was considerably enhanced by troponin I in0.05-0.08 M KC1 at pH 8.0-9.0, where the onset of superprecipitation of actomyosin was accerelated by troponin I. Electron microscopic observations revealed that the troponin I-F-actin complex was in the shape of a bundle consisting of a number of pairs of F-actin filaments (twin structure) as unit.The double stranded structure of F-actin in the twin structure was verified by optical diffraction and filtering examinations. The both end of the bundle were sharply vertical to the long axis of the bundle. When tropomyosin was added to F-actin solution before the addition of troponin I, the 400 Å periodicity was occasionally detected. Thus it is concluded that troponin I can be bound not only to tropomyosin but also directly to F-actin.View full abstractDownload PDF (19402K) -
Izumi OHKAWA, Makoto KAGEYAMA, Fujio EGAMI1973 Volume 73 Issue 2 Pages 281-289
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSPyocin S2, a bacteriocin, was purified from the mitomycin C-induced lysate of Pseudomonas aeruginosa strain M47. The preparation was chromatographically homogeneous and sodium dodesyl sulfate (SDS) disc gel electrophoresis showed a single staining band. The conditions to keep its activity stable were defined. Pyocin S2 appears to be a simple protein: it contains neither detectable amount of carbohydrate nor phosphorus. Its amino acid composition was analyzed. Its molecular weight was determined to be approximately 72, 000 from its mobility on SDS disc gel electrophoresis or 78, 000 from its sedimentation coefficient of 4.2 S combined with the elution profile of Sephadex G-200 chromatography. The frictional ratio was calculated to be 1.52.
The killing action of pyocin S2 on the sensitive cells showed a single hit process.View full abstractDownload PDF (4336K) -
Yoshiko MATSUDA, Yasuhiro KURODA, Keiko KOBAYASHI, Nobuhiko KATUNUMA1973 Volume 73 Issue 2 Pages 291-298
Published: February 25, 1973
Released on J-STAGE: June 07, 2011
JOURNAL FREE ACCESSThe mechanisms regulating serine, glycine, and glutamine metabolism were compared in ureotelic animals and uricotelic animals. The activities of five enzymes (glutaminase [EC 3. 5. 1. 2], glutamine synthetase [EC 6. 3. 1. 2], serine transhydroxymethylase [EC 2. 1. 2. 1], 5-phosphoribosylpyrophosphate amidotransferase [EC 2.4. 2. 14], and L-serine dehydratase [EC 4. 2. 1. 13]) in the livers of rats and chickens on 10 and 70% casein diets were compared. The glutaminase activity in chicken liver was less than one tenth of that in rat liver, and showed strong product inhibition by glutamate. Glutaminase activity in rat liver was induced by a high protein diet while that in chicken liver was not. A high protein diet induced glutamine synthetase in chicken liver, but not in rat liver. When a constant amount of ammonia was injected into chickens intraperitoneally, the concentration of uric acid in the serum was found to be proportional to the activity of glutamine synthetase in the liver. Phosphoribosylpyrophosphate amidotransferase activity in chicken liver was 10 fold that in rat liver, and was induced by a high protein diet while that in rat liver was not.
As already known, L-serine dehydratase activity was scarcely detectable in chicken liver, while that in rat showed high activity and was induced by a high protein diet. Serine transhydroxymethylase activity in chicken liver was high and was induced by a high protein diet while that in rat liver was low and was not induced by a high protein diet. Formation of 14C-carbon dioxide from 14C-glycine was active in chicken liver and was increased about 7-fold by a high protein diet. Its formation in rat liver was less than half that in chicken liver and was increased only 3 fold by a high protein diet. Experiments using 15N-ammonia showed that the nitrogen of inorganic ammonia was incorporated into position 7 of uric acid via glycine in chicken liver.View full abstractDownload PDF (1331K) -
I. The Effect of Fat and Its Components on Glycerokinase Activity in Rat LiverKaichi KIDA, Katsusuke KOBAYASHI, Hiroshi KIMURA, Yasumi YUGARI1973 Volume 73 Issue 2 Pages 299-306
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSThe effects of feeding rats with fat and its components, glycerol and fatty acids, on rat liver glycerokinase [EC 2.7.1.30] were investigated with the following results.
(1) Feeding of a high fat diet resulted in an increased liver glycerokinase activity. The enzyme activity obtained was proportional to the fat content of the diet.(2) The components of fat responsible for the enhanced activity were “fatty acids, ” not glycerol.
(3) Among the fatty acids tested, unsaturated fatty acids such as oleic and linoleic acids enhanced liver glycerokinase activity, whereas saturated fatty acids such as palmitic and stearic acids did not.
Based on these results, the regulatory role of unsaturated fatty acids in the metabolic relation between fat and sugar in the liver was discussed.View full abstractDownload PDF (1111K) -
Kiyoshi IKEDA, Kozo HAMAGUCHI1973 Volume 73 Issue 2 Pages 307-322
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSInteractions of divalent cations, Mn2+, Co2+, and Ni2+ with hen egg-white lysozyme [EC 3.2.1.17] were studied by the measurement of the aromatic circular dichroic (CD) spectrum. The binding constants determined by use of the decrease in the negative ellipticity of the tryptophyl CD band at 305mμ, which reflects interaction between Trp 108 and the carboxylate ion of Glu 35, were 75, 85, and 50M-1 for Mn2+, Co2+, and Ni2+, respectively, at pH7.0-7.3. On the basis of the changes in the tryptophyl CD maxima at 305 and 295mμ, it was also found that the metal ions can interact with lysozyme bound with a substrate, glycol chitin, or inhibitors, N-acetylglucosamine (NAG), di-NAG, and tri-NAG. The binding constant of Mn2+ to lysozyme was increased by a factor of about two when di-NAG or tri-NAG had been bound with lysozyme. The binding constant of Mn2+ to NAG-bound lysozyme was, however, only slightly larger than that for free lysozyme. The binding constants of di-and tri-NAG to Mn2+-bound lysozyme were also about two times as large as the constant in the absence of Mn2+, while the binding constant of NAG in the presence of Mn2+ was almost the same as that in its absence. In the case of Co2+, the binding of the saccharides was suggested to be independent of the binding of the metal ion. The enzymic activity of lysozyme toward glycol chitin in the presence of these metal ions was well explained in terms of these binding data.View full abstractDownload PDF (2408K) -
Location of Tyrosyl and Tryptophyl Residues by Solvent Perturbation Difference SpectraMasaru SOGAMI, Shigenori OGURA1973 Volume 73 Issue 2 Pages 323-334
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSIsomerization reactions of charcoal-defatted bovine plasma albumin (BPA) in 0.10 M KCl, 0.02 M NaClO4, and 0.05 M NaClO4 were studied in the acidic pH region using solvent perturbation difference spectra and optical rotation measurements. A twostep opening of BPA crevices to large perturbant, one corresponding to the N-F or N-F1 transition and the other to the acid-expansion, was observed.View full abstractDownload PDF (1872K) -
IV. Purification and Properties of the β-Xylosidase Produced by Aspergillus niger van TIEGHEMShigeyuki TAKENISHI, Yoshio TSUJISAKA, Juichiro FUKUMOTO1973 Volume 73 Issue 2 Pages 335-343
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSβ-Xylosidase [EC 3.2.1.37] of Aspergillus niger van TIEGHEM was extracted from a culture on wheat-bran koji and purified by a series of the procedures including salting out with ammonium sulfate, DEAE-Sephadex column chromatography, gel-filtration on Sephadex G-150, and column electrophoresis. The purified β-xylosidase was practically free from xylanase [EC 3.2.1.8] and all other glycosidase activities tested.
The purified enzyme was most active at pH values between 3 and 4 and stable over the range of pH's 4 to 7 at 30°C for 24hr and below 70°C for 15 min. It showed a glycosyltransferase activity, and synthesized xylooligosaccharides from xylobiose and xylotriose.View full abstractDownload PDF (5604K) -
I. Partial Purification and Characterization of RNA Polymerase IIShunji NATORI, Kazuyuki TAKEUCHI, Den'ichi MIZUNO1973 Volume 73 Issue 2 Pages 345-351
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSA DNA dependent RNA polymerase [EC 2. 7. 7. 6] was partially purified from Ehrlich ascites tumor cells. The enzyme requires Mn2+ and is inhibited by α-amanitin, a specific inhibitor of nucleoplasmic RNA polymerase. The enzyme can transcribe Ehrlich DNA and various other template DNA's including calf thymus DNA, rat ascitic hepatoma DNA, and poly dAT.
The molecular size of the enzyme is about 400, 000 daltons. The possibility of the presence of factors regulating the enzyme in vivo is discussed.View full abstractDownload PDF (1227K) -
XII. Some Properties of Flavodoxin from Desulfovibrio vulgarisKenji IRIE, Kunihiko KOBAYASHI, Miya KOBAYASHI, Makoto ISHIMOTO1973 Volume 73 Issue 2 Pages 353-366
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESS1) Flavodoxin was purified from Desulfovibrio vulgaris. The prosthetic group was identified as FMN, and amino acid content was determined. It contained one histidine and 9 cysteine (cystine), but no methionine. Molecular weight was estimated as 15, 700 from amino acid content and 15, 000 from the results of electrophoresis on sodium dodecylsulfate-polyacrylamide gel. A higher value of 25, 000 was obtained from Sephadex gel filtration.
2) The flavodoxin was found to be available as a substrate of spinach ferredoxin-NADP reductase [EC 1. 6. 99.4]. Reduction of sulfite to sulfide with hydrogen as well as phosphoroclastic cleavage of pyruvate to acetylphosphate with evolution of hydrogen and carbon dioxide in the cell-free extract of the bacteria was shown to be dependent on the presence of both cytochrome c3 and flavodoxin. The flavodoxin did not link to the solubilized hydrogenase from Desulfovibrio and was reduced by the hydrogenase system only in the presence of cytochrome c3.
3) The effect of mercurials and other reagents on dissociation of FMN from the flavodoxin was observed by measurement of fluorescence. The change in the intensity of fluorescence was followed in the course of titration of flavodoxin with phenylmercuric acetate.
4) Binding of FMN to apoflavodoxin obtained by the treatment with phenylmercuric acetate was examined. The reconstituted preparation was active as a substrate of ferredoxin-NADP reductase and stimulated phosphoroclastic cleavage of pyruvate in the extract.View full abstractDownload PDF (3974K) -
Makoto SASAKI, Shigeki IIDA, Takashi MURACHI1973 Volume 73 Issue 2 Pages 367-375
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSThe mechanism of inhibition of enzymatic activity of stem bromelain [EC 3. 4. 4. 24] by its specific antibody from rabbit sera has been studied. In the system of bromelain and its antibody, the caseinolytic activity was almost completely inhibited with enough amount of anti-bromelain antibody. The maximum number of antibody attached to one molecule of antigen was calculated to be 5 by extrapolation to the extreme excess of antibody. The ratio of antibody to antigen in the complex, whose caseinolytic activity disappeared almost completely was between 2.7 and 3.0. However, when a small molecular weight substrate as α-N-benzoyl-L-arginine ethyl ester was used the residual activity of the antigen-antibody complex was much higher than that on casein and no complete inhibition was obtained. Modification of specific amino acid residues, lysine and tyrosine by maleylation and acetylation respectively, showed considerable decreases in the amount of precipitate at each maximum point: 71% decrease by maleylation, and 50% by acetylation, whereas the enzymatic activities of the modified samples were not much altered, the activities being 81% after maleylation and 108% after acetylation. Alkylation of cysteine residue in the active site gave almost complete loss of enzymatic activity with complete preservation of antigenicity. These data suggest that the inhibition of stem bromelain by its antibody does not involve direct combination of antibody to the active site of the enzyme, but seems due to the two ways of steric hindrance according to the molecular size of the substrates. When a small molecular weight substrate was used the enzymatic activity was inhibited only by the antibodies attached to the parts of antigen molecule very close to its active site, while a large molecular weight substrate being used, steric hindrance based on the lattice formation by the antigenantibody complex plays also important role as an additional inhibition.View full abstractDownload PDF (1473K) -
Stem and Fruit BromelainsShigeki IIDA, Makoto SASAKI, Shoshi OTA1973 Volume 73 Issue 2 Pages 377-386
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSThe existence of immunological cross-reaction of stem bromelain [EC 3.4.4.24] with antiserum to fruit bromelain [EC 3.4.4.24] and of fruit bromelain with antiserum to stem bromelain was detected by gel diffusion and immunoelectrophoretic technique. This cross-reaction made it possible to attempt subsequent separation of four species of antibodies from respective antisera by means of cross-reacting enzyme immunoadsorbents. These antibody preparations comprise two species of common antibodies from anti-stem and anti-fruit bromelain antisera and two species of antibodies specific to each enzyme. The various species of antibodies were characterized and compared on the basis of their capacity to inhibit the enzymatic activity of stem and fruit bromelains and their relative reactivity in the precipitin reaction. The antibodies directed toward the specific regions of the respective enzymes exhibited pronounced precipitin reaction with the homologous enzymes, and in that case a concomitant inhibition was observed only when a substrate of large molecule was used. On the other hand, the antibody populations common to both enzymes showed to some extent a suppressed immunoprecipitation with either homologous or heterologous enzyme, inhibiting effectively the enzymatic activities without any distinction of molecular size of the substrate used.
Comparison of the inhibitory capacity and the immunological reactivity of the various antibody species led to the conclusion that the common regions of the two enzymes are mainly located in the proximity of the active site of the enzymes.View full abstractDownload PDF (9411K) -
Susumu ANDO, Tamio YAMAKAWA1973 Volume 73 Issue 2 Pages 387-396
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSGlycolipids were extracted from group-A erythrocytes with diethyl ether: methanol (1: 1), then chloroform: methanol (1: 1), and finally chloroform: methanol (1: 9). By silicic acid column chromatography, after hitherto well characterized Globoside I (A-30-I), several minor glycolipids (A-30-II, -III, A-40-VI, hematoside, A-60-III) were obtained, which were purified by repeating column chromatography. Hematoside and A-60-III contained sialic acid and were extracted effectively by chloroform: methanol (1: 9).
A-30-II was of the same carbohydrate composition with Globoside I (A-30-I), which consisted of glucose, galactose, and galactosamine (molar ratio, 1: 2: 1). The sugar moiety of A-30-III was composed of glucose, galactose, and glucosamine (1: 2: 1), the sequence of which was preliminarily determined to be Gal→GlcNAc→Gal→Glc. A-40-VI contained glucose, galactose, galactosamine, glucosamine, and fucose (1: 2: 1: 1: 1). A-60-III was composed of glucose, galactose, glucosamine, and N-acetylneuraminic acid (1: 2: 1: 1), and a glycolipid of the same structure as usual hematoside or GM3 was also obtained.
Treatment of A-60-III by neuraminidase [EC 3. 2. 1. 18] released N-acetylneuraminic acid and the glycolipid remained was identical with A-30-III on a TLC.
A-40-VI inhibited 4 units of anti-A hemagglutinin with amount of 0.02 μg/0.1ml, indicating that the material is the most purified group-A antigen on erythrocytes hitherto reported.View full abstractDownload PDF (10551K) -
Tohru NAKAZAWA, Kouichi ASAMI, Hiroko SUZUKI, Osami YUKAWA1973 Volume 73 Issue 2 Pages 397-406
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSActivities of energy conservation system in rat liver mitochondria during development were examined before and after birth. Both the respiratory control and the energy-dependent H+-efflux were extremely lower in fetal mitochondria than in adult ones. The high concentration of exogenous ADP or ATP+Mg2+ increased the energy conservation activity in fetal mitochondria, with the increase in the AdN contents. AdN contents in mitochondria increased in parallel with the respiratory control after birth, but the Pi content changed only slightly at birth. The atractyloside-sensitive uptake of [14C] AdN into mitochondria was low in fetus and increased after birth. The uptake of ATP was dependent on the concentration of exogenous ATP.View full abstractDownload PDF (1597K) -
Sueo MATSUMURA, Tsuneo OMURA1973 Volume 73 Issue 2 Pages 407-416
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSThe half-lives of messenger RNA's for liver microsomal NADPH-cytochrome c reductase and cytochrome b5 were measured with normal and phenobarbital-treated rats. After an intraperitoneal administration of actinomycin D, 4, 5-3H-L-lysine was injected intravenously to the animals at varying time points to measure the rates of incorporation of the radioactive amino acid into these microsomal enzymes. The radioactivity of lysine in the intracellular free amino acid pool of liver was also measured, and it was found that the time course of appearance and disappearance of injected radioactive lysine in the pool was greatly affected by treatment of the animals with actinomycin D. When the radioactivities of the enzymes purified from the actinomycin-treated animals were corrected for the observed changes in the radioactivity of lysine in the intracellular free amino acid pool, their corrected specific radioactivities gave exponential decay curves which were regarded to represent the decay of messenger RNA's for these enzymes in liver. The half-lives of messenger RNA's for the two enzymes were both about 3 hr.
The decrease in the rates of synthesis of these enzymes in actinomycin-treated rats was significantly slowed when phenobarbital had previously been injected to the animals. However, this effect of phenobarbital was not specific to NADPH-cytochrome c reductase whose rate of synthesis was selectively stimulated by the drug. The apparent phenobarbital-induced stabilization of messenger RNA's for microsomal enzymes does not seem to be directly correlated with the selective induction of a few particular microsomal enzymes by the drug.View full abstractDownload PDF (1902K) -
I. Tryptic Peptides of Protein NiTomotaka SHINODA1973 Volume 73 Issue 2 Pages 417-431
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSTo determine the amino acid sequence of the human κ Bence-Jones Protein Ni, the protein was completely reduced and aminoethylated and then digested with trypsin [EC 3. 4. 4. 4]. Twenty-five major tryptic peptides were isolated by chromatography on Dowex 1-×2. The complete sequences of 18 of these peptides were determined. These comprised the entire variable NH2-terminal half and 7 peptides from the COOH-terminal half of the protein. Only the amino acid compositions and partial sequences of the other peptides were determined. Only one of the tryptic peptides in the NH2-terminal half of the molecule had the same sequence as that reported for the corresponding region of other human κ chains, whereas all the tryptic peptides covering the sequence from residue 108 to the COOH-terminus corresponded exactly to the sequences reported for the constant region of human κ chains.View full abstractDownload PDF (2199K) -
II. Chymotryptic Peptides and Sequence of Protein NiTomotaka SHINODA1973 Volume 73 Issue 2 Pages 433-446
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSThe primary structure of the variable region of the human κ Bence-Jones Protein Ni was determined by sequence analysis of the tryptic and chymotryptic peptides: the primary structure of the constant region was deduced by partial sequence analysis of the peptides and by their homologies. Thirty-three chymotryptic peptides, covering all but two of the total of 218 residues, were isolated from the S-carboxymethylated protein. Complete or partial sequence analyses of the peptides covering all but two residues of the NH2-terminal half of the molecule were made to obtain overlaps for the tryptic peptides recovered from this region. For the remaining peptides covering the COOH-terminus only the partial sequences or the amino acid compositions and the end groups were determined. From these results all the tryptic peptides could be arranged in order. No evidence was obtained for variation in the sequence of the last 107 residues, but the variable region differed from others previously reported in 24 to 53 residues.View full abstractDownload PDF (2132K) -
Yoshio YAMAKAWA, Tsuneo OKUYAMA1973 Volume 73 Issue 2 Pages 447-454
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSTaka-amylase was extracted from Takadiastase with calcium acetate and purified by Sephadex G-75 and DEAE-Sephadex column chromatographies. The amino acid composition and glucosamine content of the crystallized preparation were very similar to those of Taka-amylase A (TAA)[EC 3. 2. 1. 1], but the neutral sugar content was less. The preparation was provisionally named Taka-amylase A0C. Four components with amylase activity were separated from this crystalline Taka-amylase A0C by disc electrophoresis. Two major components, named A0C-1 and A0C-2, were separated from crystalline Taka-amylase A0C by DEAE-Sephadex column chromatography. These components appeared homogeneous on disc electrophoresis. They had similar amino acid compositions, both had N-terminal alanine and a molecular weight of 51, 000, but A0C-1 had a higher content of neutral sugar than A0C-2. The molecular weights of two minor components, A0C-3 and A0C-4 were 110, 000 and 200, 000, respectively.View full abstractDownload PDF (1315K) -
Kunio YAGI, Mayumi OSAMURA, Nobuko OHISHI1973 Volume 73 Issue 2 Pages 455-457
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSDownload PDF (501K) -
Noriko ANDO1973 Volume 73 Issue 2 Pages 459-461
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSDownload PDF (476K) -
Tamaichi ASHIDA, Nobuo TANAKA, Takashi YAMANE, Tomitake TSUKIHARA, Mas ...1973 Volume 73 Issue 2 Pages 463-465
Published: February 25, 1973
Released on J-STAGE: January 25, 2011
JOURNAL FREE ACCESSDownload PDF (2716K)
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