巻号一覧
63 巻, 1 号
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YONG-CHI LIN, NOBUO TANAKA1968 年 63 巻 1 号 p. 1-7
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリーThe poly UC- or poly UG-directed incorporation of phenylalanine, proline, leucine, serine and valine into polypeptide was inhibited by bottromycin A2 and its hydrazide. The formation of a ternary complex between the ribosomes, poly C and prolyl-sRNA was not significantly affected by bottromycin A2 or its hydrazide. Bottromycin did not cause significant peptide release nor inhibit puromycin-dependent release of polyproline from ribosomes. The activity of bottromycin A2 was reversed by a high concentration of ribosomes but not by that of the 105, 000×g supernatant or poly C. The binding of chloramphenicol to the ribosomes was not antagonized by bottromycin A2 or its hydrazide. The inhibitory activity of bottromycin A2 seemed to be reversible, because the effect of the antibiotic on the ribosomes or supernatant was lost on dialysis.抄録全体を表示PDF形式でダウンロード (428K) -
HYOGO SINOHARA, YOSHIKO ASANO1968 年 63 巻 1 号 p. 8-13
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリー1. D-Glucosamine-1-14C, N-acetyl-D-glucosamine-1-14C, D-glucosaminic acid-1-14C, and D-glucose-1-14C were administered to larvae of the silkworm, and the amounts of the isotope in expired CO2 and silk proteins were determined.
2. After 4 hours, CO2 exhaled from feeding larvae contained 14.5, 8.7, 2.5, and 1.5% of isotope injected as glucose, glucosaminic acid, glucosamine, and N-acetylglucosamine, respectively. In spinning larvae, considerable changes were observed in the rate of oxidation of these amino sugars.
3. Fibroin and sericin were isolated from cocoons and hydrolyzed to determine the specific activities of amino acids, hexosamines, and hexoses. All the compounds investigated were efficiently incorporated into silk proteins not only as hexosamines and hexoses, but also as serine, alanine, glycine, aspartic, and glutamic acids. No radioactivity was associated with other amino acids. The labeling patterns obtained with glucosamine and N-acetylglucosamine were similar to that with glucose, suggesting that these sugars were mainly metabolized by the same pathway. Experiments with glucosaminic acid revealed a considerable difference in the relative distribution of isotope among amino acids, suggesting that the catabolism of this compound is different from that of glucose.
4. These results indicate that hexosamine metabolism in the silkworm differs markedly from that in mammals and microorganisms.抄録全体を表示PDF形式でダウンロード (471K) -
EI-ICHI SOEDA, JUN-ICHIRO MUKAI, SATORU AKUNE1968 年 63 巻 1 号 p. 14-19
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリーThe silkworm nuclease [EC class 3. 1. 4] had no definite base specificity but shown only a slight base preference. This enzyme attacked RNA in an endonucleolytic manner. The rate of enzyme reaction was influenced by the chain length rather than the nature of bases in substrate and was very slow when the substrate reached a critical chain length. Digestion of RNA with an excess of the nuclease produced exclusively mono-, di- and trinucleotides. Of the mononucleotide fraction, 5'-guanylic acid predominated over the other three.
This enzyme may be a useful reagent in the structural studies on RNA.抄録全体を表示PDF形式でダウンロード (392K) -
YASUKO MURAKAMI, KATASHI MAKINO1968 年 63 巻 1 号 p. 20-24
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリーDecarboxylation of 5-hydroxykynurenine to 5-hydroxykynuramine and the subsequent oxidation of the latter to form 4, 6-dihydroxyquinoline were shown to occur in mouse liver homogenate.
The identification of 5-hydroxykynuramine was carried out by paper chromatography, enzymatic conversion to 4, 6-dihydroxyquinoline and spectrophotometric analysis. 4, 6-Dihydroxyquinoline was identified by paper chromatography and fluorometric analysis.抄録全体を表示PDF形式でダウンロード (304K) -
YOSHIO YAMASAKI, KEISUKE MURAKAMI, MASACHIKA IRIE, TYUNOSIN UKITA1968 年 63 巻 1 号 p. 25-32
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリー1. A ribonuclease (RNase P) was isolated from porcine pancreas. The enzyme was purified by means of ammonium sulfate fractionation, phenol extraction, DEAE-, Phospho- and CM-cellulose column chromatography about 850 fold.
2. Purified RNase P was homogeneous in centrifugation (S=2.10). The specific activity of RNase P was about 70-85% of RNase A [EC 2. 7. 7. 16, ribonucleate pyrimidinenucleotido-2'-transferase (cyclizing)] under the conditions used for estimation.
3. RNase P was very similar to that of RNase A in respect to molecular weight, heat stability and the activity dependence on ionic strength.
4. The substrate specificity of the enzyme was qualitatively the same as that of RNase A and the enzyme was inhibited by Cu++, Zn++ and Hg++ as in the case of RNase A.
5. The pH optima of RNase P were at pH 7.5 (RNA as substrate) and 6.0 (cytidine-2', 3'-cyclic phosphate (C-cyclic-P) as substrate).
6. The isoelectric point and elution positions from the columns (CM- and Phospho-cellulose) of RNase P indicated that this ribonuclease was less basic than RNase A.抄録全体を表示PDF形式でダウンロード (1290K) -
TAKASHI YAMAUCHI, JIRO KOYAMA, IKUO YAMASHINA1968 年 63 巻 1 号 p. 33-40
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリー1. Human α1-AGP*** produces antibodies in rabbit sera which vary in their ability to precipitate the antigen, from practically zero up to about 80% in the antibody excess region.
2. From the supernatant of immune reactions, soluble antigenantibody complexes were isolated by means of electrophoresis and gel filtration.
3. Antigenic properties of polymorphic variants of α1-AGP were investigated using the antibodies with different precipitating capacities. However, α1-AGP and its subfractions as well as sialic acid-free α1-AGP were found to be indistinguishable from each other in their immunological properties.抄録全体を表示PDF形式でダウンロード (1211K) -
XV. Ethyl-morpholinylpropyl-carbodiimide for Modification of Carboxyl Groups in ProteinsHIROO HORINISHI, KAZUYASU NAKAYA, AKIKO TANI, KAZUO SHIBATA1968 年 63 巻 1 号 p. 41-50
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリーA water soluble carbodiimide, EMPC, was applied in the presence of glycine ethylester to acetylglycine and proteins, and was found to be a reagent suitable for discrimination of different states of carboxyl groups in the tertiary structures of proteins. One of the total two carboxyl groups in the bacitracin molecule, at least three of the six carboxyl groups in bovine insulin and six of the eleven carboxyl groups in egg white lysozyme [EC 3. 2. 1. 17] were modified by the treatment with 40 mM EMPC for 3 hours at room temperature and no further reaction took place at higher EMPC concentrations. All of these carboxyl groups were modified when the proteins had been denatured with guanidine. The results obtained in these measurements were compared with previous modification data in view of the state discrimination of carboxyl groups.抄録全体を表示PDF形式でダウンロード (746K) -
SATOSHI NAKAMURA, SUMIKO FUJIKI1968 年 63 巻 1 号 p. 51-58
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリー1. The molecular weights of the glucose oxidases [EC 1. 1. 3. 4] of Penicillium amagasakiense and Aspergillus niger were measured by YPHANTIS' method and found to be 150, 000 and 152, 000, respectively.
2. The carbohydrate and amino acid compositions in these enzymes were compared. Both enzymes contained the same kinds of carbohydrate, that is, mannose, glucose and hexosamine. There was more mannose and hexosamine and less glucose in the Aspergillus-enzyme than in the Penicillium-enzyme. There was more histidine, arginine and tyrosine and less lysine and phenylalanine in the Aspergillus-enzyme than in the Penicillium-enzyme.
3. The two enzymes were titrated with D-glucose under anaerobic conditions, but no intermediate, such as a semiquinone or charge-transfer type compound, appeared in the equilibrium state between the oxidized and reduced forms. On titration with dithionite, an intermediate having a semiquinone character was formed with the Aspergillus-enzyme, but not with the Penicillium-enzyme. The absorption spectrum of the inter-mediate of the Aspergillus-enzyme was obtained on the basis of the value of the molecular extinction coefficient at each wavelength. The latter was calculated from the equilibrium relationship between the oxidized, semiquinone and reduced forms of the FAD moiety.抄録全体を表示PDF形式でダウンロード (593K) -
MICHIYUKI YAMADA, KIYOSHI KURAHASHI1968 年 63 巻 1 号 p. 59-69
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリーA phenylalanine racemase [EC 5. 1. 1 group] was found to be present in the cell-free extract of Bacillus brevis NAGANO, which produces gramicidin S. This enzyme was partially purified by ultracentrifugation, ammonium sulfate precipitation, and calcium phosphate gel adsorption and elution. The enzyme showed an absolute requirement of adenosine triphosphate, inorganic pyrophosphate, and magnesium or manganese ion for its activity. Thiol compounds, such as dithiothreitol and 2-mercaptoethanol stimulated the enzyme activity. The pH optimum of the reaction was between 8.2 and 8.5. The rate of the formation of L-phenylalanine from D-phenylalanine catalyzed by this enzyme lower than that of n-phenylalanine from L-phenylalanine. When the reaction reached an apparent equilibrium, L-phenylalanine and D-phenylalanine were found to be in the ratio of 3 to 7.
This racemase activity appeared abruptly at the middle of the logarithmic phase of growth, and increased towards the end of the logarithmic phase of growth. When the cell entered the stationary phase, the enzyme activity disappeared rapidly. The similar change of the rate of gramicidin S production and of the activities of gramicidin S synthesizing system and L- and D-phenylalanine-activating enzyme was also observed.抄録全体を表示PDF形式でダウンロード (796K) -
II. Further Studies on the Properties of the Two HemoglobinsMASATOMO YOSHIOKA, KINZABURO HAMADA, TARO OKAZAKI, AKIHIKO KAJITA, RYO ...1968 年 63 巻 1 号 p. 70-76
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリー1. Chromatographic separation of the two hemoglobins (HbE1 and HbE2) in the blood of the eel, Anguilla japonica, into their constituent subunits was studied. It was found that both hemoglobins were composed of four polypeptide chains, each having the valyl residue as the amino terminal amino acid. Analyses of the carboxyl terminal amino acids and of the tryptic peptides, however, indicated that there was no polypeptide chain common to the two hemoglobins. The amino acid compositions of the subunits of the two hemoglobins were also studied.
2. Studies on the oxygen equilibria of the two hemoglobins revealed that they differed from each other with respect to their oxygen affinities, heme-heme interaction and the Bohr effect. HbE1 was half-saturated with oxygen at 2.1 mmHg, while HbE2 was half-saturated at 14mmHg, at pH 7.0 and 20°C. The n value of Hill's equation was 2.4 for HbE1, and 1.0-1.8 for HbE2 varying with the pH. A Bohr effect was observed with HbE2, but not with HbE1.
The oxygen equilibria of a hemolysate, a solution of HbE1 together with HbE2, and an erythrocyte suspension were also measured and compared with each other.抄録全体を表示PDF形式でダウンロード (479K) -
YUKINORI KAWAHARA, EIICHI MISAKA, KAZUO NAKANISHI1968 年 63 巻 1 号 p. 77-82
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリーLipoamide dehydrogenase [EC 1. 6. 4. 3] was purified 310-fold and obtained in crystalline form from a yeast, Candida krusei. Purification and crystallization of the enzyme was accomplished by the following steps; disruption of cells by autolysis with ethyl acetate, precipitation by ammonium sulfate, absorption and elution on calcium phosphate gel, chromatography on TEAE-cellulose at pH 7.0 and crystallization by dialysis. This enzyme showed very similar properties to the enzymes from various sources.抄録全体を表示PDF形式でダウンロード (1063K) -
I. Glycolytic Intermediates and Nucleotides in Guinea Pig Exudate GranulocytesSHIGEKI MINAKAMI1968 年 63 巻 1 号 p. 83-88
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリー1. Glycolytic intermediates and adenine nucleotides in polymorphonuclear granulocytes from guinea pig peritoneal exudate were analyzed. Glucose 6-phosphate, fructose 6-phosphate, fructose 1, 6-diphosphate, dihydroxyacetone phosphate, glyceraldehyde 3-phosphate, 3-phosphoglycerate, 2-phosphoglycerate, phosphoenolpyruvate, pyruvate, ATP, ADP and NAD+ were measured enzymatically using a fluorometer. Nucleotides were also analyzed by column chromatography on Dowex 1-formate. Peritoneal exudate was deproteinized either immediately after removal from the body to obtain an aerobic steady state pattern or after incubating with cyanide to get an anaerobic steady state pattern.
2. Mass action ratios of glycolytic reactions in leucocytes were calculated from the above data and rate-limiting steps in the cellular glycolysis were discussed. Hexokinase [EC 2. 7. 1. 1], phosphofructokinase [EC 2. 7. 1. 11] and pyruvate kinase [EC 2. 7. 1.40] were found to be major rate-limiting steps in leucocytes as in the case of erythrocytes.
3. Following changes were observed when leucocytes were incubated in presence of cyanide: 1) decrease in hexose monophosphates, 2) increase in fructose diphosphate and triose phosphates, 3) decrease in monophosphoglycerates and phosphoenolpyruvate and 4) slight decrease in ATP and increase of ADP.抄録全体を表示PDF形式でダウンロード (401K) -
III. The Reaction MechanismHIDEO INOUE, FUJIO SUZUKI, HIROOAKI TANIOKA, YOSHIRO TAKEDA1968 年 63 巻 1 号 p. 89-100
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリーThe mode of action of ATP citrate lyase [EC 4. 1. 3. 8] was investigated using a single homogeneous preparation from rat liver. First, the enzyme was incubated with labelled citrate and after incubation the protein portion was isolated through a column of Sephadex G-50. It was found that an enzyme-citrate complex is formed in the presence of both ATP and Mg++. When this enzyme-citrate complex was isolated and then incubated with CoA, the formation of acetyl-CoA was demonstrated in the absence of added ATP.
When 8-14C-ATP and λ-32P-ATP were separately incubated with the enzyme, radioactivity associated with protein was obtained only with λ-32P-ATP, but not with 8-14C-ATP. In accord with this, a rapid ATP-ADP exchange, but not an ATP-P1 exchange, was observed. These two reactions were solely dependent on the presence of Mg++. Therefore, the reaction involves the formation of an enzyme-phosphate complex.
When the enzyme-phosphate complex, labelled with 32P, was isolated through a Sephadex G-50 column and then incubated with labelled citrate in the presence of CoA, oxaloacetate was formed quantitatively on the basis of bound phosphate in the absence of added ATP. Mg++ was unnecessary in this conversion. Bound phosphate was almost completely released from the enzyme on addition of citrate. It was also found that the over-all reaction of ATP citrate lyase is reversible, though it is stronger in the cleavage direction. From these findings, the reaction mechanism of ATP citrate lyase can be summarized as follows: Mg++
1. Enzyme+ATPMg++_??_Enzyme-phosphate+ADP
2. Enzyme-phosphate+citrate_??_Enzyme-citrate+Pi
3. Enzyme-citrate+CoA_??_Acetyl-CoA+oxaloacetate+enzyme
The properties of the reactive intermediates were also described.抄録全体を表示PDF形式でダウンロード (971K) -
YUTARO HAYASHI, YUJI TONOMURA1968 年 63 巻 1 号 p. 101-118
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリーGlycerol-treated muscle fiber bundles were prepared mainly by storage in 50 percent glycerol at -10°C for about 3 months. Their ATPase [EC 3. 6. 1. 3] activity was measured in the presence of 50mm KCl, 2 to 3.35mM MgCl2 and 20mM Tris-maleate buffer at pH 7.0 and 25°C during an isometric contraction induced by ATP. Pyruvate kinase [EC 2. 7. 1. 40] was coupled with myofibrillar ATPase, and the activity was measured by determining the amount of pyruvate liberated. The process of liberation of pyruvate from fiber bundles was linear with time in all the measurements.
The ATPase activity of fiber bundles of about 300μ in diameter and 2.2 to 2.6μ in sarcomere length was independent of the concentration of phosphoenol pyruvate between 0.5 and 3.0mM and the concentration of pyruvate kinase between 5 and 32μg per ml, in the presence of 0.6mM ATP. Furthermore, in the presence of 1.0mM phosphoenol pyruvate and 65μg pyruvate kinase per ml, the activity was scarcely affected by an increase in concentration of ATP from 0.6 to 5.0mM. The ATPase activity of fiber bundles of 250 to 310μ in diameter and 4.2 to 5.3μ in sarcomere length was independent of the concentration of ATP between 0.22 and 0.77mM or of that of phosphoenol pyruvate between 0.5 and 1.0mM, if the amounts of other constituents were sufficient. The activity of fiber bundles of 240 to 310μ in diameter and 0.8 to 0.9μ in sarcomere length was not affected by increase in the concentration of pyruvate kinase from 25 to 250μg per ml or that of ATP from 2.35 to 5.0mM. The activity in 0.6mM ATP was almost equal to that in 2.35mM ATP. Thus, it was concluded that the diffusion of ATP and phosphoenol pyruvate was not the limiting factor in the hydrolysis of ATP, if fiber bundles of less than 300μ in diameter were used in the presence of at least 0.6mM ATP and 1.0mM phosphoenol pyruvate.
The fiber bundles showed the maximum activity of 100 pmoles pyruvate per minute per g of protein at sarcomere lengths of 2.0 to 2.5μ. As the sarcomere length increased beyond 2.5 p, the activity decreased and reached about 30 pmoles pyruvate per minute per g of protein at sarcomere lengths of more than 4μ. In the region where the sarcomere length was less than 2.0μ, the ATPase activity decreased as the sarcomere length decreased, and, at a sarcomere length of 1μ, it showed a quarter to a third of the maximum activity. These results suggested that both ATPase activity and tension development are coupled with the movement of myosin filaments past actin filaments. The maximum activity observed at sarcomere lengths of 2.0 to 2.5μ was only one fourth of the activity of the actomyosin type of isolated actomyosin.
The dependence on Ca++ concentration of the ATPase of glyceroltreated muscle fiber bundles, which had been stored for about 3 months, was investigated at fixed sarcomere lengths of 2.0 to 2.6μ. When Ca++ was removed by EGTA, the activity was about 48 μmoles py ruvate per minute per g of protein, and in the presence of 1 mat Ca++ it was 116 μmoles pyruvate per minute per g of protein. The concentration of Ca++ for the half maximum activation was about 0.04 μM. In the presence of 1 to 2mM EGTA, the activity of fiber bundles stored for 3 to 13 days was only 17 μmoles pyruvate per minute per g of protein, and that of those stored for about 9 months was 126 μmoles pyruvate per minute per g of protein.抄録全体を表示PDF形式でダウンロード (2472K) -
XII. Determination of the Carboxyl-terminal Structure of Clupeine and Salmine Using Enzymatic ProceduresTOHRU TOBITA, MAKOTO YAMASAKI, TOSHIO ANDO1968 年 63 巻 1 号 p. 119-126
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリー1. The amino acid sequences of the carboxyl-end of clupeine-1 (a protamine prepared from Clupea pallasii), clupeine-2 (Clupea harengus) and salmine (Oncorhynchus keta) were investigated by applying alternate action of highly purified porcine pancreatic carboxypeptidases B and A [EC 3. 4. 2. 2 and 3. 4. 2. 1]. It is concluded that only the stepwise release of amino acids from the carboxyl-end of the protein along the peptide chain has occurred.
2. From clupeine-1 and -2, 4 moles of arginine followed by 0.9-1 mole of alanine, together with trace amounts of glycine, serine, and threonine, were released per mole of protamine by the procedure. The results are interpreted to mean that both clupeine specimens mainly have the same carboxyl-terminal sequence of …Ala•Arg4•OH.
3. When the remaining clupeine-1 “core” was subjected to the second cycle of alternate enzymatic action, arginine (3.4 moles per mole of the “core”), and serine (0.67 mole) followed by valine (0.75 mole) were released. The results and informations obtained from the sequence study of tryptic digests of a clupeine specimen, suggest that a molecular species with the following carboxyl-terminal sequence, …(Arg)•Val•Ser•Arg4•Ala•Arg4•OH, composes 70-80% of clupeine-1.
4. In the case of salmine, only 3 moles of arginine were released by the alternate digestion. When hydrazinolysis-DNP-procedure was applied to the salmine “core”, net 0.69 mole of di-DNP-ornithine per mole of the “core” was recovered. It is suggested that the salmine specimen has the carboxyl-terminal sequence of …(Pro_??_)•Arg•Arg3•OH.
5. Analyzing the way of tryptic hydrolysis of the ester bond of clupeine-1 and salmine methyl esters by using hydroxamate-ferric ion colorimetry, it is concluded that, in both protamines, the arginine content at the carboxyl-end amounts to at least 95%.抄録全体を表示PDF形式でダウンロード (569K) -
KYUJI OHTA, MIKI GOTO1968 年 63 巻 1 号 p. 127-129
発行日: 1968/01/25
公開日: 2008/11/18
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NAOMI AZUMA1968 年 63 巻 1 号 p. 130-132
発行日: 1968/01/25
公開日: 2008/11/18
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IKUYA YANO, YOSHIYA FURUKAWA, MASAMICHI KUSUNOSE1968 年 63 巻 1 号 p. 133-135
発行日: 1968/01/25
公開日: 2008/11/18
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GENJI MATSUDA, TETSUO MAITA, MASATO YAMAGUCHI, HISAHIRO OTA, MASAHARU ...1968 年 63 巻 1 号 p. 136-137
発行日: 1968/01/25
公開日: 2008/11/18
ジャーナル フリーPDF形式でダウンロード (143K)
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