巻号一覧
83 巻, 4 号
選択された号の論文の40件中1～40を表示しています
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I. Isolation and Preliminary CharacterizationYukinobu HINO, Akira ASANO, Ryo SATO, Satoru SHIMIZU1978 年 83 巻 4 号 p. 909-923
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーA method is described for the isolation of morphologically well-preserved Golgi apparatus from rat liver. The method is essentially the same as that of Morré et al. (Morré, D. J., Hamilton, R. L., Mollenhauser, H. H., Mahley, R. W., Cunningham, W. P., Cheetham, R. D., & Lequire, V. S. (1970) J. Cell Biol. 44, 484-491) except that mild cell disruption is achieved by means of a stainless-steel sieve. The average recoveries of protein and galactosyltransferase in the isolated fraction are about 6mg from 10g of perfused liver and about 35% from the homogenate, respectively. The preparation is virtually free from succinate-cytochrome c reductase, but contains significant activities of NADH- and NADPH-cytochrome c reductase, glucose-6-phosphatase, acid phosphatase, and 5'-nucleotidase. The Golgi fraction as well as its vesicular fragments is homogeneous upon isopycnic centrifugation in both sucrose and dextran density gradients. Their buoyant densities in sucrose are significantly higher than those in dextran, indicating that both forms of the organelle are closed systems which are impermeable to macromolecules. The galactosyltransferase activity of a freshly prepared Golgi fraction, measured with ovalbumin as galactosyl acceptor, is activated 26-fold by the addition of Triton X-100, whereas those of homogenized, sonicated, and aged preparations are only activated 2- to 4-fold.抄録全体を表示PDF形式でダウンロード (2666K) -
II. Further Characterization of Isolated Golgi FractionYukinobu HINO, Akira ASANO, Ryo SATO1978 年 83 巻 4 号 p. 925-934
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーAlthough the preparation of rat liver Golgi apparatus isolated by our method contains appreciable activities of NADH- and NADPH-cytochrome c reductases and glucose-6-phosphatase, these enzymes as well as thiamine pyrophosphatase of the extensively fragmented Golgi fraction are partitioned in aqueous polymer two-phase systems quite differently from those associated with microsomes. Similarly, the partition patterns of acid phosphatase and 5'-nucleotidase of the Golgi fragments differ from those of homogenized lysosomes and plasma membrane, respectively. It is concluded that most, if not all, of these marker enzymes in the Golgi fraction cannot be ascribed to contamination by the non-Golgi organelles. In sucrose density gradient centrifugation the NADH- and NADPH-cytochrome c reductase activities of the Golgi fraction behave identically with galactosyltransferase but differently from the reductase activities of microsomes, again indicating that the reductases are inherently associated with the Golgi apparatus. NADPH-cytochrome c reductase of the Golgi preparation is immunologically identical with that of microsomes. The marker enzymes mentioned above and galactosyltransferase behave differently from one another when the Golgi fragments are subjected to partitioning in aqueous polymer two-phase systems, suggesting that these enzymes are not uniformly distributed in the Golgi apparatus structure.抄録全体を表示PDF形式でダウンロード (762K) -
III. Subfractionation of Fragmented Golgi Apparatus by Counter-Current DistributionYukinobu HINO, Akira ASANO, Ryo SATO1978 年 83 巻 4 号 p. 935-942
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーVesicular fragments of Golgi apparatus, smooth- and rough-surfaced microsomes from rat liver are differently partitioned in aqueous polymer two-phase systems consisting of dextran, polyethylene glycol, and sodium phosphate buffer. At a given polymer concentration, the amount of material partitioned in the top phase increases in the following order: rough microsomes<smooth microsomes<Golgi fragments. Counter-current distribution of Golgi fragments in the system consisting of 6.8% (w/w) dextran T500 and 6.8% polyethylene glycol 4, 000 results in the separation of the fragments into three fractions; i.e. Fractions I, II, and III. NADH- and NADPH-cytochrome c reductase activities are detected almost exclusively in Fraction I, whereas the activities of galactosyltransferase, acid phosphatase, 5'-nucleotidase, and thiamine pyrophosphatase are maximal in Fraction III and minimal in Fraction I. The distribution of these enzymes suggests that Fraction I is similar to, though not identical with, microsomes, Fraction III resembles plasma membrane and lysosomes, and Fraction II is between the two. It is concluded that NADH- and NADPH-cytochrome c reductases are localized in a restricted region of the Golgi structure and that intra-Golgi differentiation seems to proceed in a discontinuous manner.抄録全体を表示PDF形式でダウンロード (631K) -
Kimie YASUO, Nobuo TSUCHIDA, Katsuji HORI1978 年 83 巻 4 号 p. 943-951
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーA conditional mutant, referred to as RepR43, was isolated from Escherichia coli W2252 by N-methyl-N'-nitro-N-nitroso-guanidine mutagenesis.
Although RepR43 does not permit growth of RNA phage β at the restrictive temperature, 43°C, cell growth and synthesis of macromolecules such as RNA and protein continue at a somewhat reduced rate.
Several lines of evidence indicate that a RepR43 function is indispensable for normal phage RNA replication. In addition, this function appears to be involved in the maintenance of the perpetuated phage genome.
The addition of 10% sucrose to the medium at the restrictive temperature resulted in the production of the phage, suggesting that the mutant cell might have an altered membrane organization which interferes with normal viral replication.抄録全体を表示PDF形式でダウンロード (656K) -
Nobuko SENO, Etsuko SEKIZUKA1978 年 83 巻 4 号 p. 953-956
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーThree different types of chondroitin polysulfate-peptide, chondroitin sulfate D-peptide, chondroitin sulfate E-peptide, and chondroitin sulfate K-peptide, all contained xylose, galactose, and serine in a molar ratio of about 1:2:1. After treatment with alkali in the presence of NaBH4 and PdCl2, they produced alanine and xylitol in amounts equivalent to the decrease in the amount of serine. Consequently, it was proved that these chondroitin polysulfates are all linked to peptides by 0-glycosidic bonds between xylose and serine, as in chondroitin sulfates A and C. It is suggested that the carbohydrate-peptide linkage regions have the same structure in all the chondroitin sulfates, regardless of differences in the structure of the polysaccharide chains, such as the position of sulfate groups and the degree of sulfation.抄録全体を表示PDF形式でダウンロード (260K) -
Chang Gie KANG, Michio MUGURUMA, Tatsumi ITO, Toshiyuki FUKAZAWA1978 年 83 巻 4 号 p. 957-960
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーSingle sarcomeres were prepared from fresh rabbit myofibrils by digestion with a calciumactivated factor (CAF). The rabbit single sarcomere has functional properties quite similar to those of single sarcomeres obtained from chicken muscle by a usual method. Thus it was found that the single sarcomeres obtained by CAF digestion were useful as a muscle model, though they were not completely intact.抄録全体を表示PDF形式でダウンロード (1317K) -
Takayoshi IIO, Koshin MIHASHI, Hiroshi KONDO1978 年 83 巻 4 号 p. 961-969
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーThe kinetics of conformational change of troponin-C (TN-C) induced by binding or removal of calcium ion were studied in the presence or absence of magnesium ion by measuring the fluorescence of tyrosyl residues by stopped-flow spectrofluorometry. The result was analyzed in terms of first-order kinetics. Two phases were observed both in pCa-up and in pCa-down experiments. The dependence of the rate constants on pCa was explained by a simple mecha-nism as follows;
The dissociation constants of calcium bound to TN-C, K and K', calculated from the experimentally determined rate constants were K=3.16×10-7M, K'=1.58×10-6M in the absence of magnesium ion, and K=K'=1×10-6M in the presence of 2 mM MgCl2.抄録全体を表示PDF形式でダウンロード (545K) -
Ken-ichi KODAIRA, Akira TAKETO1978 年 83 巻 4 号 p. 971-976
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーHost functions required for replication of progeny double-stranded DNA of bacteriophage G4 were examined by using metabolic inhibitors and Escherichia coli dna mutants. In dna+ bacteria, synthesis of the progeny replicative form (RF) was relatively resistant to 30μg/ml of chloramphenicol, but considerably sensitive to 200μg/ml of rifampicin. The RF replication was severely inhibited by 50μg/ml of mitomycin C, 50μg/ml of nalidixic acid, or 200μg/ml of novobiocin. At 41°C, synthesis of G4 progeny RF was distinctly affected in a dnaC(D) mutant and in a dnaG host. The progeny RF replication was prevented at 42°C in a dnaE strain as well as in a dnaB mutant. In a dnaZ strain, the synthetic rate of the progeny RF was markedly reduced at 42°C. At 43°C, the rate of G4 progeny RF synthesis was reduced even in dna+ or dnaA bacteria, but significant amounts of the progeny RF were still synthesized in these hosts at the high temperature. In addition to five dna gene products, host rep function was essential for the RF replication.抄録全体を表示PDF形式でダウンロード (433K) -
Motonori YAMAGUCHI, Yuji TONOMURA1978 年 83 巻 4 号 p. 977-987
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーNa+, K+-dependent ATPase [EC 3.6.1.3] was partially purified from porcine kidney by the method of Lane et al. with slight modifications. The properties of partial reactions of the enzyme were compared with those of the NaI-treated enzyme from bovine brain, which had been used in our previous studies. The following results were obtained: (1) no difference was observed in the elementary processes which constituted these two ATPase reactions, (2) but the affinity of the porcine kidney enzyme for K+ ions was about four times that of the porcine brain enzyme.
On the basis of these results, the amount of ADP bound to the enzyme from porcine kidney was measured during ATP hydrolysis in the presence of 4mm MgCl2, 100mM NaCl, and 100mM Tris-HCl at pH 8.5, using sufficient amounts of creatine kinase (CK) [EC 2.7.3.2] and creatine phosphate (CP) to regenerate [14C]ATP from free [14C]ADP. The amounts of ADP bound to the enzyme were determined by separating the nucleotides using TLC after stopping the reaction with TCA. The following results were obtained.
1. At 15°C and in the absence of CP all the phosphorylated intermediate (EP) was ADP-insensitive. On the other hand, in the presence of CK and CP the amount of ADP bound to the Na+, K+-dependent ATPase in the absence of K+ ions was about one-third of the amount of EP.
2. The amount of ADP bound to the enzyme decreased slightly with an increase in the KCl concentration to 0.5mM, but remained almost constant in the KCl concentration range of 1-100mM. On the other hand, the amount of EP decreased markedly with increasing KCl concentration. Thus, the amount of EP was much smaller than that of ADP bound to the enzyme in 1-100mM KCl. The rate of the ATPase in the steady state, V0, was almost proportional to the amount of ADP bound to the enzyme in the range of 1-100mM KCl.
3. However, at 1°C about half of the EP was ADP-sensitive even in the presence of a high concentration of MgCl2, and the amount of ADP-insensitive EP was equal to that of bound ADP in the NaCl concentration range of 10-100mM.抄録全体を表示PDF形式でダウンロード (823K) -
Yoshinobu NAKANISHI, Den'ichi MIZUNO, Shunji NATORI1978 年 83 巻 4 号 p. 989-994
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーThe molecular size and poly-A content of RNA synthesized in isolated nuclei of Ehrlich ascites tumor cells were measured. KCl was found to be essential for synthesis of high molecular weight RNA: when 0.4M KCl was added to the reaction mixture, the average molecular size of the RNA formed was 14S; without KCl the average molecular size was 5S.
A significant amount of poly-A sequences was found in RNA synthesized in the presence of α-amanitin, suggesting that RNA polymerase I and/or III may synthesized some RNA containing poly-A in isolated nuclei.抄録全体を表示PDF形式でダウンロード (462K) -
Tsukasa SUGANO, Kohei SUDA, Masakazu SHIMADA, Nozomu OSHINO1978 年 83 巻 4 号 p. 995-1007
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーThe viability of hemoglobin-free perfused rat liver was examined with respect to several liver functions and to the intactness of subcellular structures under electron microscopic observation. Provided that rat livers were perfused with the oxygenated buffer solution at a flow rate between 3 and 3.5ml/min per g of liver, all the biochemical parameters measured in the perfused liver system, i.e. the rates of glucose, pyruvate, and lactate production, the rate of oxygen consumption and the tissue contents of adenine nucleotides, were similar to those observed with perfusion systems containing erythrocytes or albumin. The perfused liver showed a sensitive response to norepinephrine, involving a reduction of pyridine nucleotides and enhancements of glucose production and oxygen consumption.
On electron microscopic examination, changes in hepatic-structure indicative of hypoxic injury particularly vacuolar degeneration and mitochondrial swelling, were not detected in the liver after 70min of perfusion; the fact that the fine structure of the hepatocyte was preserved in all parts of the organ confirmed that the supply of oxygen to the perfused liver was sufficient under the conditions employed.
From viewpoint of the generally accepted criteria for the viability of perfused liver, therefore, the results confirmed that the perfusion of liver with a hemoglobin- and albumin-free medium is a convenient and reliable tool for biochemical investigation of the reactions occurring in whole liver.抄録全体を表示PDF形式でダウンロード (3217K) -
I. Polysaccharide from E. coli 36MAkira KAMEI, Noriko TAKEUCHI, Shuzo AKASHI, Koshiro KAGABE1978 年 83 巻 4 号 p. 1009-1017
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーAn extracellular polysaccharide was isolated from culture broth of Escherichia coli 36M, and fractionated on a column of Sephadex G-150 into two fractions; the high molecular weight portion (85%. of the total polysaccharide) contained pyruvic acid, and showed a positive immune reaction with anti-Ps-I-serum obtained from a rabbit. The low molecular weight portion (15% of the total polysaccharide) showed a negative immune reaction.
The methylation, Smith's degradation, partial acid hydrolysis and methanolysis of the higher molecular weight polysaccharide revealed a repeating structure as follows:抄録全体を表示PDF形式でダウンロード (570K) -
Daikichi FUKUSHIMA, Yong H. KIM, Takashi IYANAGI, Shigeru OAE1978 年 83 巻 4 号 p. 1019-1027
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーBoth rabbit liver microsomes and reconstituted system with purified cytochrome P-450 and cofactors enzymatically oxidized o-dithiane (1, 2-dithiane), 3-methyl-o-dithiane, thiane and 2-methylthiane to the corresponding mono-oxygenated products; sulfides or disulfides were oxidized to the corresponding sulfoxides or thiolsulfinates, while thiolsulfinate was oxidized to thiolsulfonate. The reconstituted systems required oxygen and NADPH and were not affected by the catalase which decomposes H2O2, or by 1, 4-diazabicyclo-[2, 2, 2]octane (DABCO), which is a good quencher of ringlet oxygen.
The differences in the binding of substrates such as sulfides and disulfides with the enzyme system are discussed in connection with differences in the spectra of the substrates in the reconstituted system with pure cytochrome P-450. A correlation was found between the rates of oxidation of the substrates and the rates of oxidation of NADPH.抄録全体を表示PDF形式でダウンロード (699K) -
Akira MURASUGI, Shosuke TAKEMURA1978 年 83 巻 4 号 p. 1029-1038
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーTwo species of 32P-labelled leucine tRNA were highly purified from Candida (Torulopsis) utilis by successive column chromatographies. The purified major species of leucine tRNA 1 was completely digested with ribonuclease T1 [EC 3.1.4.8] and with pancreatic ribonuclease A [EC 3.1.4.22]. The resulting fragments were fractionated, and their nucleotide sequences were determined according to Barrell (1). The results of analyses of the two ribonuclease digests were consistent with each other, and indicated that this tRNA is composed of 85 nucleotide residues, including 14 modified nucleotides. A tentative total sequence has been derived on the basis of several features in the cloverleaf structure for tRNA.抄録全体を表示PDF形式でダウンロード (596K) -
Yoshiie OKADA, Tsuneo OMURA1978 年 83 巻 4 号 p. 1039-1048
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリー1. The turnovers of NADH-cytochrome b5 reductase, cytochrome b5, and NADPH-cytochrome c reductase in rat liver were determined by two separate methods including a double-label method using 14C- and 3H-leucines. Although NADH-cytochrome b5 reductase and cytochrome b5 are functionally and spatially associated in the microsomal membrane, their turnover rates were definitely different, confirming the independent turnover of microsomal membrane proteins.
2. The NADH-cytochrome b5 reductase of mitochondrial outer membrane had the same turnover rate as the corresponding enzyme of microsomes. As the composition and functions of mitochondrial outer membrane are significantly different from those of microsomes, the identical turnover rates of the reductases associated with these two membranes suggest that the degradation of membrane-bound molecules of the reductase is catalyzed by a factor extrinsic to the membranes.
3. After a single intravenous injection of 3H-leucine into rats, the labeled NADH-cytochrome b5 reductase molecules appeared first in the membrane of rough microsomes and subsequently in smooth microsomes. The appearance of radioactive reductase molecules in the outer membrane of mitochondria lagged behind that in smooth microsomes. At about 3h after the injection, the labeled reductase molecules were almost equally distributed among rough and smooth endoplasmic reticula and mitochondrial outer membrane. Possible mechanisms for the intracellular translocation of the reductase from the site of synthesis to different membrane systems are discussed.抄録全体を表示PDF形式でダウンロード (775K) -
Shin-ichi KUWAHARA, Yoshiie OKADA, Tsuneo OMURA1978 年 83 巻 4 号 p. 1049-1059
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーTo confirm the molecular identity of the NADH-cytochrome b5 reductase of mitochondrial outer membrane with the corresponding enzyme of microsomes, the properties of mitochondria) and microsomal reductases were examined and compared. The NADH-cytochrome b5 reductase associated with rat liver mitochondria was easily solubilized by digestion with cathepsin D, exactly as the corresponding microsomal reductase was. The cytochrome b5-reducing activities of microsomal and mitochondrial reductases were both stimulated by about 80% upon solubilization from the membranes, whereas such stimulation was not observed when potassium ferricyanide was used as the electron acceptor. Some physicochemical properties and reaction kinetics of the solubilized microsomal and mitochondrial reductases were examined, and no significant differences were found between them.
The rabbit antibody prepared against purified microsomal NADH-cytochrome b5 reductase inhibited the rotenone-insensitive NADH-cytochrome c reductase activities of microsomes and mitochondria to the same extent. The immunological identity of microsomal and mitochondrial reductases was confirmed by the Ouchterlony double diffusion test in agar gel and by quantitative immunoprecipitation tests using the cathepsin-solubilized enzyme preparations. The NADH-cytochrome b5 reductase molecules of mitochondria) outer membrane seem to be located on.the outside surface of the membrane, since the antibody against the reductase inhibited the rotenone-insensitive NADH-cytochrome c reductase activity of intact mitochondria as well as that of hypotonic-treated mitochondria. The solubilization of the reductase from intact mitochondria by cathepsin D also supported this conclusion. It is thus concluded that the same reductase molecules are present on the cytoplasmic surfaces of endoplasmic reticulum and mitochondrial outer membrane in liver cells.抄録全体を表示PDF形式でダウンロード (1428K) -
Haruo SUZUKI1978 年 83 巻 4 号 p. 1061-1064
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーThe effect of concentration of rabbit reticulocyte rRNAs on the translation of rabbit globin mRNAs was studied by using a rabbit reticulocyte lysate system. Globin synthesis was studied using L-[U-14C]leucine.
Both the 18S and 28S rRNA inhibited globin synthesis. The 18S rRNA inhibited the synthesis of α-globin chain more than that of β chain, but the 28S rRNA inhibited the synthesis of α- and β-chains almost equally.
Nascent chains prelabelled with [14C]Leu or f[35S]Met were released at various concentrations of rRNAs. Release of the nascent chains was not inhibited at various concentrations of rRNAs. The ratios [14C]α/[14C]β and [35S]α/[35S]β in the released chains were almost constant at various concentrations of rRNAs. It therefore appears that the inhibition of globin synthesis by these rRNAs was at the step of initiation.抄録全体を表示PDF形式でダウンロード (297K) -
Shohei MAEKAWA, Hikoichi SAKAI1978 年 83 巻 4 号 p. 1065-1075
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーTetrahymena tubulin was purified from the cell extract using DEAE-Sephadex A-50 ionexchanger and ammonium sulfate precipitation. About 2.2% of the total protein in the 20, 000×g supernatant was recovered as DEAE-Sephadex-purified tubulin fraction. Applying the temperature-dependent polymerization-depolymerization method to this fraction in the presence of Tetrahymena outer fibers as a seed, almost pure tubulin was obtained.
Tetrahymena tubulin dimer showed different behavior on SDS-polyacrylamide gels from porcine brain tubulin, and showed very low affinity for colchicine, amounting to about onetwentieth of the binding to porcine brain tubulin.
The tubulin fraction failed to polymerize into microtubules by itself. Addition of a small amount of the ciliary outer fiber fragment induced polymerization, as demonstrated by viscometric measurements, but the reconstituted microtubules were very unstable in the absence of glycerol. Microtubule-depolymerizing agents such as Ca2+ ions, low temperature, or colchicine all inhibited in vitro polymerization. Although Tetrahymena tubulin purified by the polymerization-depolymerization method could copolymerize with porcine brain microtubules, the DEAE-Sephadex-purified tubulin fraction suppressed the initial rate of porcine brain microtubule assembly in vitro.
There seemed to be no differences between cytoplasmic tubulin and outer fiber tubulin in colchicine binding activity or SDS-gel electrophoretic behavior, or between the fine structure of both reconstituted microtubules observed by electron microscopy.抄録全体を表示PDF形式でダウンロード (2694K) -
X. Heat of Formation of the Intermediate in the Bioluminescent Reaction Studied by Stopped-Flow CalorimetryTakao NAKAMURA1978 年 83 巻 4 号 p. 1077-1083
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーThe heat production in the reaction of luciferase-FMNH, complex with O2 in the absence of aldehyde was measured by stopped-flow calorimetry. ΔH of the reaction, luciferase-FMNH2+O2→intermediate X1, is -1.3×102 kJ•mol-1 and the calculated ΔS for the reaction is -180 J•mol-1•K-1 at 20°C. The heat production in the bioluminescent reaction was also measured in the presence of a saturating concentration of aldehyde, and it was estimated that 43 and 79 of the C10 and C13 aldehydes, respectively, bound with the intermediate X1 are converted to carboxylic acid yielding energy for photon emission.抄録全体を表示PDF形式でダウンロード (489K) -
Kenichi NAKANO, Sadao WAKABAYASHI, Toshiharu HASE, Hiroshi MATSUBARA, ...1978 年 83 巻 4 号 p. 1085-1094
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーThe amino acid sequence around the pyridoxal 5'-phosphate binding site in potato phosphorylase was determined in order to compare it with those in phosphorylases from other sources having different regulatory properties. The potato enzyme was reduced by NaBH4 in the presence of urea, carboxymethylated, and digested with chymotrypsin and trypsin. Pyridoxyl peptides were isolated by the differential procedure using paper electrophoresis or DEAE-cellulose column chromatography. In Edman degradation of these peptides, pyridoxyllysine was identified as the phenylthiohydantoin derivative of pyridoxyllysine using a combination of thin-layer chromatography and the Pauli reaction. The sequence around pyridoxyllysine, comprising 57 amino acid residues, was determined except for a region with 6 amino acid residues. The pyridoxal 5'-phosphate binding site in potato phosphorylase showed a high homology with those of the rabbit muscle and yeast enzymes. This finding suggests that the cofactor should be directly related to the essential process of phosphorylase action.抄録全体を表示PDF形式でダウンロード (637K) -
Evidence that Structural Genes for the Two Proteins are DifferentShigeyuki ICHIHARA, Shoji MIZUSHIMA1978 年 83 巻 4 号 p. 1095-1100
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーOuter membrane proteins O-8 and O-9 have been highly purified from a strain of Escherichia coli K-12 by Sephadex G-200 and DEAE-cellulose chromatographies. The amino acid compositions of the purified proteins were definitely different, although they showed marked similarities. The profiles of BrCN peptides of the two proteins were also different. None of the BrCN peptides were the same for the two proteins. Analysis of the first twelve N-terminal residues revealed that the two proteins are strikingly similar, but with differences in the third and the eleventh amino acid residues. It can be concluded that proteins O-8 and O-9 are products of different structural genes which developed by duplication of an ancestral genome followed by mutation.抄録全体を表示PDF形式でダウンロード (875K) -
Sumiko YASUE, Shizuo HANDA, Sachiko MIYAGAWA, Jun INOUE, Atsuhiko HASE ...1978 年 83 巻 4 号 p. 1101-1107
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーHematoside from dog erythrocyte membrane was previously considered to contain a mixture of N-acetyl- and N-glycolyl- neuraminic acids. However, the hematoside preparation used in the previous study was obtained from pooled blood of several dogs, and individual variation in hematoside was not examined. In this work, hematosides of erythrocytes from 31 mongrel dogs and 108 dogs of 23 breeds were examined individually by thin-layer chromatography, and the component sialic acids were analysed by gas-liquid chromatography. Individual dogs had either NAN-hematoside or NGN-hematoside: dogs with N-glycolyl-neuraminic acid also had a trace of N-acetyl-neuraminic acid, but dogs with N-acetyl-neuraminic acid had no detectable N-glycolyl-neuraminic acid. A few mongrel dogs, some Kai dogs, Kishu dogs, Japanese spaniels and most Shiba dogs had NGN-hematoside, whereas all European dogs had NAN-hematoside and no NGN-hematoside. From pedigrees of some families, inheritance of NGN-hematoside was found to be autosomal dominant. NGN-hematoside is possibly one of dog blood group substances. The sialic acid of delipidized ghost protein of dogs with NGN-hematoside was N-glycolyl-neuraminic acid, and that of dogs with NAN-hematoside was N-acetyl-neuraminic acid. The sialic acid of plasma protein was mainly N-acetyl-neuraminic acid in all dogs.抄録全体を表示PDF形式でダウンロード (1139K) -
Nakao ARIGA, Hiroshi HATANAKA, Jun NAGAI, Hirohiko KATSUKI1978 年 83 巻 4 号 p. 1109-1116
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーIn order to identify the methyl acceptor for the methylation of sterol side-chains in ergosterol biosynthesis, Saccharolnyces cerevisiae (wild type) was grown in the presence and absence of ethionine which was expected to be an inhibitor of the methylation. Gas-liquid chromatographic analyses of the sterols in the cells grown in the absence of ethionine showed that ergosterol was the most abundant sterol. On the other hand, a sterol, named sterol Z, accounted for more than 50% of the total sterols in the cells grown in the presence of ethionine. As a result of experiments to raise the yield of sterol Z, the best concentration of DL-ethionine for the production was found to be 1.0mM. The use of the methionine-less mutant was less effective for the production of sterol Z. Sterol Z was isolated by repeated TLC and was identified as zymosterol from its melting point, GLC and mass spectrometry.
The role of zymosterol and other sterols as the methyl-acceptor sterol in ergosterol biosynthesis is also discussed.抄録全体を表示PDF形式でダウンロード (556K) -
I. Chemical CompositionSeiichi KAWASHIMA, Toshio ANDO1978 年 83 巻 4 号 p. 1117-1123
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーThe chemical composition of deoxyribonucleoproteins from herring sperm nuclei was analyzed and the results are summarized as follows.
1. Chemical analysis of nuclear proteins and nucleic acids revealed that arginine/P molar ratio in herring sperm nuclei is unity but the ratio of arginine residues in protamine to phosphorus in the total DNA is 0.86.
2. The deoxyribonucleoproteins were isolated and their composition showed that about 14% of the total DNA in herring sperm nuclei is free from protamine and is bound with nonprotamine proteins in the weight ratio of nonprotamine proteins to DNA of 0.25-0.30. The remaining 86% of the total DNA is combined mainly with protamine and a small amount of nonprotamine proteins; the weight ratios of protamine and nonprotamine proteins to DNA are 0.75 and 0.08, respectively. In the latter complex, the molar ratio of arginine residues in protamine to phosphorus in DNA is unity.抄録全体を表示PDF形式でダウンロード (476K) -
Kiyofumi MARUYAMA, Nakao ARIGA, Mitsushige TSUDA, Kazuo DEGUCHI1978 年 83 巻 4 号 p. 1125-1134
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーα-Hydroxy-γ-Carboxymuconic ε-semialdehyde (HCMS) dehydrogenase was purified about 180-fold with a yield of 22% from a cell-free extract of Pseudomonas ochraceae. The enzyme was adsorbed tightly on blue dextran, possibly at its coenzyme binding sites. This finding made possible a simple purification step by molecular exclusion chromatography on Sephadex G-200. The enzyme adsorbed on blue dextran was eluted in the void volume and was well separated from contaminating proteins which were eluted at various positions. The purified enzyme migrated as a single band on disc gel electrophoresis. The molecular weight of the enzyme as determined by gel filtration on Sephadex G-200 was 67, 000. Polyacrylamide gel electrophoresis of the enzyme after sodium dodecyl sulfate denaturation gave a single band at a position corresponding to a molecular weight of 35, 000. The enzyme showed a simple protein absorption in the UV region and no significant absorption in the visible region. The enzyme activity was not affected by various metal ions, metal chelating reagents or reducing reagents, but was strongly inhibited by various sulfhydryl reagents. Both NAD and NADP were found to be efficient coenzymes. Various aldehyde derivatives such as acetaldehyde, propionaldehyde, n-butyraldehyde, iso-butyraldehyde, succinaldehyde, crotonaldehyde, chloral hydrate, glyoxylic acid, DL-glyceraldehyde, benzaldehyde, salicylaldehyde, glucose, and protocatechualdehyde did not serve as substrates. The stoichiometry of HCMS consumption and coenzyme reduction was approximately 1:1. The reaction product, which appeared to be α-hydroxy-γ-carboxymuconic acid, showed an absorption maximum at 311nm at pH 8.0.抄録全体を表示PDF形式でダウンロード (772K) -
Yoshiaki SONE, Akira MISAKI1978 年 83 巻 4 号 p. 1135-1144
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーβ-D-Mannosidase [EC 3.2.1.25] and β-N-acetyl-D-hexosaminidase [EC 3.2.1.30] were purified approximately 500- and 200-fold, respectively, from the cell extract of Tremella fuciformis. Both glycosidases showed single protein bands in disc gel electrophoresis and the molecular weights of β-D-mannosidase and β-N-acetyl-D-hexosaminidase were about 140, 000 and 125, 000, respectively, as estimated by Sephadex gel exclusion chromatography. The substrate specificities and kinetics of the two enzymes were tested with p-nitrophenyl glycosides and related oligosaccharides. The β-D-mannosidase hydrolyzed p-nitrophenyl β-D-mannoside, with Km 2.1mM and Vmax 0.21 μmol per min per mg protein. 4-O-β-D-Mannosyl-D-mannose was readily hydrolyzed, but β-(1→4)-linked mannotriose and mannotetraose were hydrolyzed much more slowly. Unlike other known β-D-mannosidases, its activity was not inhibited by D-manno-γ-lactone, but was strongly inhibited by p-chloromercuribenzoate. The β-D-mannosidase tended to be inactivated in the presence of oxygen but was reactivated by dithiothreitol. The β-N-acetyl-D-hexosaminidase hydrolyzed p-nitrophenyl β-N-acetyl-D-glucosaminide, with Km 0.31mM and Vmax 4.6 μmol per min per mg protein. It was active on N, N'-diacetylchitobiose and higher saccharides (DP up to 5) and liberated N-acetyl-D-glucosamine. The glycosidase preparation was also slightly active on p-nitrophenyl β-N-acetyl-D-galactosaminide (Km 0.19mM, and Vmax 0.6 μmol per min per mg protein). Both β-D-mannosidase and β-N-acetyl-D-hexosaminidase had an optimum pH of 5.0. Inhibition of both glycosidases by various metal ions was tested and their stabilities were investigated.抄録全体を表示PDF形式でダウンロード (707K) -
Kunio FUJIWARA, Daisuke TSURU1978 年 83 巻 4 号 p. 1145-1149
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーL-Pyroglutamyl derivatives of p-nitroaniline and 7-amino-4-methylcoumarin were synthesized as new sensitive substrates for pyrrolidonyl peptidase (pyrrolidonecarboxylyl peptidase) from Bacillus amyloliquefaciens. Their hydrolyses could be followed by conventional colorimetric and fluorometric procedures; i.e., in terms of the increase in absorbance at 410 nm caused by the liberation of p-nitroaniline and the emission at 440nm after excitation at 370nm depending on the liberation of 7-amino-4-methylcoumarin. Values of Km were estimated to be 0.69mM for anilide substrate and 0.33mM for methylcoumarin substrate in the pyrrolidonyl peptidase reaction at pH 8.0. The methylcoumarin compound was about one thousand fold more sensitive than the anilide substrate.抄録全体を表示PDF形式でダウンロード (322K) -
III. Two Ferredoxins from Pokeweed, Phytolacca americana; Isolation and CharacterizationShinichi TAKAICHI, Kazuo MAKINO, Akio IWASAKI, Koichi SUZUKI1978 年 83 巻 4 号 p. 1151-1158
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーTwo ferredoxins were isolated from pokeweed, Phytolacca americana, and purified by ammonium sulfate fractionation (60-100% saturation), Sephadex G-50 and DEAF-cellulose column chromatographies, and preparative polyacrylamide disc gel electrophoresis at pH 9.3. The faster running ferredoxin on the gel electrophoresis was designated as Fd I and the slower one as Fd II of P. americana.
The absorption spectra of Fd I and Fd II were very similar to those of Scenedesmus and spinach ferredoxins, respectively, exhibiting absorption maxima at 275 (276 for Fd II), 330, 422, and 465nm. A422/A275 was 0.70 for Fd I and A422/A276 was 0.48 for Fd II. Amino acid compositions were as follows: Fd I; Lys4, His1, Arg1, Asp13, Thr11, Ser6, Glu13, Pro5, Gly7, Ala9, Cys(5), Val9, Ile4, Leu6, Tyr3, Phe3, 100 residues in total: Fd II; Lys3, His1, Arg1, Trp1, Asp9, Thr11, Ser7, G1u14, Pro5, Gly7, Ala13, Cys(5), Val7, Met1, Ile2, Leu6, Tyr4, Phe2, 99 residues in total. Two non-heme irons and two acid-labile sulfides were detected per molecule for both ferredoxins. Molecular weights as determined by Sephadex G-75 column chromatography were 11, 500±500 for both ferredoxins and those calculated from the amino acid compositions were 10, 617 and 10, 438 for Fd I and Fd II, respectively. ε422 of the oxidized form was found to be 9.85×103 for both ferredoxins.
The photochemical activities of both ferredoxins as estimated with washed, broken chloroplasts of spinach were found to be almost identical.抄録全体を表示PDF形式でダウンロード (1853K) -
Masahiko YAMAMOTO, Zensaku YOSIZAWA1978 年 83 巻 4 号 p. 1159-1164
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーDesialized ovine submaxillary major mucin was digested extensively with pronase. The digestion product was gel-filtered through Sephadex G-25 then fractionated by ion-exchange column chromatography on Hitachi custom resin No. 2614. Of the resulting fractions, the fractions containing relatively simple glycopeptides were purified by rechromatography on the same ion-exchange column, followed by preparative high-voltage paper electrophoresis. The isolated glycopeptides, A12d, A14a, A17c, and B12, were shown to be homogeneous by high-voltage paper electrophoresis at pH 1.9, 3.7, and 6.5 and by paper chromatography.
The results of chemical analysis, alkali treatment, determination of the amino acid sequence and digestion with α-N-acetylgalactosaminidase of these glycopeptides indicated their structures to be as follows: glycopeptide A12d, O-α-N-acetylgalactosaminyl-Ser-Glx-Pro-Gly; glyco-peptide A14a, O-α-N-acetylgalactosaminyl-Ser; glycopeptide A17c, O-α-N-acetylgalacto-saminyl-Ser-Gly-Gly-(O-α-N-acetylgalactosaminyl-)-Thr-Glx; glycopeptide B12, Gly-(O-α-N-acetylgalactosaminyl-)-Ser-Ala.抄録全体を表示PDF形式でダウンロード (408K) -
Showbu SATO, Kayoko NAKAZAWA1978 年 83 巻 4 号 p. 1165-1171
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーManganese-containing superoxide dismutase was isolated from an extreme thermophile, Thermos thermophilus HB8. About 150mg of the enzyme was obtained from 500g of wet cells. The enzyme was easily crystallized in octahedra from ammonium sulfate solution. The molecular weight of the enzyme was determined to be 8.2×104 and 8.4×104 by sedimentation equilibrium and gel-filtration, respectively. The enzyme contains 2 atoms of manganese per mole and consists of four subunits of identical molecular weight, about 2.1×104. The amino acid composition of the enzyme is similar to that of the superoxide dismutase of Thermus aquaticus. Proline was detected as the N-terminal amino acid. The isoelectric point was determined to be pH 6.0 by the electrofocusing method. The enzyme has maxima at 283nm and 480nm in the absorption spectrum. The CD spectrum suggests that the enzyme has a high α-helical content.抄録全体を表示PDF形式でダウンロード (1584K) -
Takao YORA, Nobuo Ul1978 年 83 巻 4 号 p. 1173-1190
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーProcedures for the group separation of bovine pituitary glycoprotein hormones, i.e., thyrotropin, lutropin, and follitropin, and for the purification and subfractionation of the first two hormones were studied. Radioimmunoassay systems specific to bovine thyrotropin and lutropin have been developed and used to evaluate these purification processes. Pituitary glycoprotein hormones were fractionally extracted with aqueous ethanol from 400g of bovine lyophilized pituitary powder by the percolation method and purified by bioaffinity chromatog, raphy on a column of concanavalin A-Sepharose; more than 500-fold purification was achieved within 4-5 days and the estimated overall yield was close to 90% of the total extractable hormones. A mixture of thyrotropin and lutropin was obtained efficiently from this glycoprotein hormone concentrate by CM-cellulose chromatography and further fractionated by DEAE-cellulose chromatography. Although most of the lutropin was separated from thyrotropin on this chromatography, thyrotropin was always accompanied by a substantial amount of lutropin. On isoelectric focusing of the thyrotropin-containing fraction thus obtained, five peaks of immunologic thyrotropin with isoelectric points at pH 7.20, 7.90, 8.32, 8.60, and 8.95 were obtained; contaminating lutropin was also distributed over this pH range. Thyrotropin components completely free of lutropin could be obtained by the use of affinity chromatography on a column of immobilized anti-bovine lutropin antibodies. On the other hand, four components of bovine lutropin, completely or almost completely free of thyrotropin, were purified by isoelectric focusing of two lutropin fractions obtained by DEAE-cellulose chromatography. Their isoelectric points were distributed over a pH range of 8.2-9.0.抄録全体を表示PDF形式でダウンロード (1457K) -
Takahide TSUCHIYA, Tetsuo KANEKO, Juichiro J. MATSUMOTO1978 年 83 巻 4 号 p. 1191-1193
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーCa2+ -sensitivity of squid muscle proteins was studied by competition tests between squid actin and carp myosin on squid myosin B and by cross combination tests between squid myosin and actin and between carp myosin and squid thin filament fraction. The results suggest that the regulatory system of the squid mantle muscle is actin-linked as in skeletal muscle rather than myosin-linked. This view is supported by the presence of troponin-like components on SDS-disc electrophoresis of squid thin filament fraction.抄録全体を表示PDF形式でダウンロード (169K) -
II. Occurrence of 3-O-Methylgalactosamine in Oyster GlycolipidToshiko MATSUBARA, Akira HAYASHI1978 年 83 巻 4 号 p. 1195-1197
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリー3-O-Methyl hexosamine was found for the first time and characterized as one of the sugar components of oyster glycolipid. The alditol acetate of this sugar was identified as 3-0-methyl-N-acetylgalactosaminitol acetate by comparing its retention time on gas chromatography and mass spectrum with those of the authentic sample synthesized in this laboratory. The new sugar, 3-0-methylgalactosamine, occupied the non-reducing terminal position of the carbohydrate moiety of the lipid and was linked to the penultimate hexose by a (1→3) bond.抄録全体を表示PDF形式でダウンロード (190K) -
Kei-ichi UEMURA, Matsuko YUZAWA, Tamotsu TAKETOMI1978 年 83 巻 4 号 p. 1199-1201
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーAntibodies against a glycolipid of guinea pig erythrocyte membranes were prepared in rabbits by immunization with guinea pig erythrocyte stroma or the purified glycolipid, gangliotriaosyl-ceramide.1 The antibodies agglutinated guinea pig erythrocytes. The specificity of antibodies could be revealed by several immunochemical methods, including inhibition of hemagglutination, immunodiffusion, agglutination of liposomes, and complement fixation. The antibodies were specific for gangliotriaosylceramide.抄録全体を表示PDF形式でダウンロード (203K) -
Masafumi YANO1978 年 83 巻 4 号 p. 1203-1204
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーSteady streaming of the solution was observed for more than one and half hours in a circular slit between two concentric cylinders where rabbit skeletal F-actin was fixed on to the wall of the slit and heavy meromyosin was in the Mg-ATP solution in the slit. The streaming only appeared when the actomyosin-ATPase activity was present in the system.抄録全体を表示PDF形式でダウンロード (149K) -
Masaaki HIROSE, Masatoshi MAKI, Hideo CHIBA1978 年 83 巻 4 号 p. 1205-1207
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーThe template activities of chromatin isolated from virgin, pregnant, lactating, and regressed rat mammary glands were compared by using exogenously added bacterial and mammalian RNA polymerases. Template activity increased during the stage of pregnancy, then decreased during the stage from early lactation to regression. It showed the highest level at early lactation when intracellular levels of rRNA and mRNA are known to be at their maximums. These results suggest that transcriptional activity in the mammary gland is regulated, at least in part, by a modulation of chromatin template activity.抄録全体を表示PDF形式でダウンロード (232K) -
Synthesis of the 3'-Pyrophosphates of Pyrimidine Nucleotides, Some Oligoribonucleotides, 5'-Diphosphonucleosidic Coenzymes and 7mG-5-ppp-5-AmJun-Ichiro MUKAI, Toshio KUKITA, Sawao MURAO, Toyokazu NISHINO1978 年 83 巻 4 号 p. 1209-1212
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーATP: nucleoside-5'-phosphate pyrophosphotransferase [EC 2.7.6.4] of Streptomyces adephospholyticus synthesizes not only 3'-pyrophosphates of 5'-purine ribomononucleotides but also those of pyrimidine mononucleotides, some short oligonucleotides, a variety of 5'-diphos-phonucleosidic coenzymes and 7mG-5'-ppp-5'-Am.抄録全体を表示PDF形式でダウンロード (230K) -
Akihiro YAMAMOTO, Ikuya YANO, Masamiki MASUI, Eiko YABUUCHI1978 年 83 巻 4 号 p. 1213-1216
発行日: 1978/04/25
公開日: 2008/11/18
ジャーナル フリーA new acidic sphingoglycolipid has been isolated from a Gram-negative, glucose-non-fermentative (obligatory aerobic) bacterium, Flavobacterium devorans ATCC 10829, by thin-layer chromatography on silica gel after mild alkaline hydrolysis of the cellular lipids. Chemical degradation studies, thin-layer chromatographic behavior, 1R and mass-spectrometric analysis of the original and reduced glycolipid with LiAIH4 revealed that the lipid contained glucuronic acid, long-chain bases, and fatty acids in a molar ratio of approximately 1:1:1. The major long-chain bases were identified by gas chromatography-mass spectrometry as dihydrosphingo-sine (d-18 : 0) and longer homologues, while the N-acyl group was exclusively 2-hydroxy myristic acid. The most probable structure of this glycolipid appeared to be a ceramide glucuronic acid (N-acyl dihydrosphingosine 1-glucuronic acid).抄録全体を表示PDF形式でダウンロード (750K) -
1978 年 83 巻 4 号 p. 1217a
発行日: 1978年
公開日: 2008/11/18
ジャーナル フリーPDF形式でダウンロード (37K) -
1978 年 83 巻 4 号 p. 1217b
発行日: 1978年
公開日: 2008/11/18
ジャーナル フリーPDF形式でダウンロード (37K)
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