巻号一覧
95 巻, 4 号
選択された号の論文の39件中1～39を表示しています
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Nobuyuki FUJITA, Tetsuya MIWA, Sumio ISHIJIMA, Katsura IZUI, Hirohiko ...1984 年 95 巻 4 号 p. 909-916
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーThe nucleotide sequence of the ppc gene, the structural gene for phosphoenolpyruvate carboxylase [EC 4. 1. 1. 31], of Eseherichia coli K-12 was determined. The gene codes for a polypeptide comprising 883 amino acid residues with a calculated molecular weight of 99, 061. The amino acid sequence deduced from the nucleotide sequence was entirely consistent with the protein chemical data obtained with the purified enzyme, including the NH2- and COOH-terminal sequences and amino acid composition. The coding region is preceded by two putative ribosome binding sites, and is followed closely by a good representative of rho-independent terminator. The codon usage in the ppc gene suggests a moderate expression of the gene. The secondary structure of the enzyme was predicted from the deduced amino acid sequence.抄録全体を表示PDF形式でダウンロード (602K) -
Kristina Nilsson EKDAHL, Pia EKMAN1984 年 95 巻 4 号 p. 917-924
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーAfter partial chromatographic purification of rat liver cell sap on DEAE-cellulose, including the removal of type M2 pyruvate kinase, different forms of type L pyruvate kinase were separated by chromatofocusing. Three fractions of pyruvate kinase activity were found, eluting at pH 5.0, 5.2, and 5.3, respectively. The first one was identified as phosphorylated and the second one as unphosphorylated pyruvate kinase. There were strong indications that the third fraction represented a proteolytically modified form of the enzyme, since it co-migrated with a form modified in vitro and had a similarly increased apparent Km for phosphoenolpyruvate. To rule out the possibility of this being a phosphorylated form of pyruvate kinase, the enzyme was incubated with a phosphoprotein phosphatase and then phosphorylated with cAMP-dependent protein kinase. The enzyme was not phosphorylated, like pyruvate kinase modified with subtilisin or calcium-activated protease. There is some evidence that a proteolytically modified pyruvate kinase exists in vivo. This enzyme form has not previously been demonstrated in cell sap, prior to exposure to proteolytic enzymes. The relative amounts of the three forms were determined in livers from starved rats and rats fed on a normal or a carbohydraterich diet.抄録全体を表示PDF形式でダウンロード (1477K) -
Kaneki KOYAMA, Minehiko YOKOYAMA, Tsuneaki KOIKE, Kenzo OHTSUKI, Nakao ...1984 年 95 巻 4 号 p. 925-935
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーA phosphate-incorporating protein has been highly purified from the cytosol of Ehrlich ascites tumor cells (EAT cells). The nitrocellulose membrane method was used to follow the progress of the purification by quantitation of the [32P] phosphorylated form of the protein. The purified protein was identified as an NDP-kinase since it exhibited NDP-kinase activity and had enzyme characteristics in common with other NDP-kinases from various mammalian cells. The purified NDP-kinase was found to have a molecular weight of approximately 76, 000 daltons. Moreover, the enzyme appears to consist of two distinct polypeptides (18, 000 and 20, 000 daltons). This enzyme contained 19 amino acids, with high levels of glycine (9.8%) and lysine (9.0%). The enzyme rapidly formed a [32P] phosphoenzyme when incubated with [γ-32P] ATP in the presence of Mg2+ (1mM) at the optimum pH of 7.5 even at low temperature (below 4°C). This phosphoenzyme is an enzyme-bound, high-energy-phosphate intermediate, because ATP was formed from it on incubation with ADP in the presence of Mg2+ (1mM). This finding suggests that the phosphoenzyme functions as an intermediate in NDP-kinase action.抄録全体を表示PDF形式でダウンロード (2864K) -
Hidefumi YOSHIOKA, Toshiyuki MIYATA, Tsuneo OMURA1984 年 95 巻 4 号 p. 937-947
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーWhen 1, 1-di (p-chlorophenyl)-2, 2-dichloroethylene (DDE) (100mg/kg body weight) was injected into rats, the benzphetamine N-demethylation and 7-ethoxycoumarin O-deethylation activities of liver microsomes increased by 7-fold and 3-fold, respectively, at 7 days after the injection, whereas the benzo(a)pyrene 3-hydroxylation activity did not increase. The content of cytochrome P-450 in the microsomes increased 2.5-fold at 7 days. By the use of the antibodies to a phenobarbital (PB)-inducible form (P-450 (PB-1)) and a 3-methylcholanthrene (MC)-inducible form (P-450 (MC-1)) of cytochrome P-450, the contents of P-450 (PB-1) and P-450 (MC-1) in the liver microsomes of DDE-treated rats were measured. The form of cytochrome P-450 immunoprecipitable with anti-P-450 (PB-1) antibodies increased by 10-fold at 7 days.
A major component of cytochrome P-450 in the liver microsomes of DDE-treated rats, which was tentatively designated P-450 (DDE), was purified. P-450 (DDE) was compared with P-450 (PB-1), and they were found to be indistinguishable by the following criteria: 1) chromatographic behavior on aminooctyl-Sepharose 4 B, hydroxyapatite, and DEAE-cellulose columns, 2) minimum molecular weight determined by SDS-polyacrylamide gel electrophoresis, 3) spectral properties, 4) immunoreactivity, 5) amino acid composition, 6) peptide mapping, 7) NH2-terminal amino acid sequence, 8) catalytic activities. We concluded that DDE and PB induce an identical form of P-450 in rat liver microsomes, although DDE is apparently very different from PB in chemical structure.抄録全体を表示PDF形式でダウンロード (2401K) -
Ken-ichirou MOROHASHI, Hidefumi YOSHIOKA, Kazuhiro SOGAWA, Yoshiaki FU ...1984 年 95 巻 4 号 p. 949-957
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーIn the preceding paper (Yoshioka, H., et al. (1984) J. Biochem. 95, 937-947), we reported that 1, 1-di (p-chlorophenyl) -2, 2-dichloro-ethylene (DDE) induced the pheno-barbital (PB)-inducible form of microsomal cytochrome P-450 (P-450 (PB-1)) in rat liver. In order to study more precisely the molecular events responsible for the induction of this particular form of cytochrome P-450 by the two chemical compounds, we determined the amounts of the mRNA coding for P-450 (PB-1) in the liver of rats given a single dose of PB or DDE. RNA was extracted from the livers of the treated rats and the determination of the specific mRNA was carried out by using the rabbit reticulocyte lysate translation system and by a dot hybridization method using cloned P-450 (PB-1) cDNA (Fujii-Kuriyama, Y., et al. (1982) Proc. Natl. Acad. Sci. U. S. 79, 2793-2797) as the probe. The amounts of P-450 (PB-1) mRNA determined by these two methods at various time points of the induction process showed good agreement. These observations further confirmed the induction of an identical form of cytochrome P-450 by DDE and PB. The maximum level of P-450 (PB-1) mRNA, which was about 8-fold higher than the control level, was attained at 20-30h and at 48-72h after the administration of PB and DDE, respectively. The mRNA level showed a rapid decrease after the peak in the liver of PB-treated rats, but the decrease was much slower with DDE-treated rats. We conclude that DDE had a more persistent inducing effect on the mRNA level than PB, although these two compounds induced an identical form of cytochrome P-450 in the liver microsomes of the animals.抄録全体を表示PDF形式でダウンロード (2478K) -
Tomoko ITOH, Yutaka UDA1984 年 95 巻 4 号 p. 959-970
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーSquid liver contains two kinds of α-N-acetylgalactosaminidases, which could be separated by gel filtration on Sephadex G-200 or by SP-Sephadex ion exchange chromatography. The two α-N-acetylgalactosaminidases, α-N-acetylgalactosaminidase I and II, were purified by procedures involving extraction, ammonium sulfate precipitation, and chromatographies on SP-Sephadex, Sephadex G-100, Sephadex G-200, DEAE-Sephadex, and Sepharose 6 B. Enzyme I was purified 1, 100-fold and enzyme II 3, 000-fold. Both enzymes appeared to be homogeneous based upon the results of disc gel electrophoresis. Enzyme I had a pH optimum of 3.0 and was heat-stable. It was inhibited by N-acetylgalactosamine and galactose. On the other hand, enzyme II had a pH optimum of 4.2 and was heat-labile. Galactose did not affect the enzyme activity. In contrast to enzyme I, which showed α-galactosidase activity even in the final preparation, enzyme II was practically free from α-galactosidase activity.抄録全体を表示PDF形式でダウンロード (1356K) -
Masanobu ARITA, Masao IWAMORI, Tetuo HIGUCHI, Yoshitaka NAGAI1984 年 95 巻 4 号 p. 971-981
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーHigh molecular weight gangliosides (GD1a, GD1b, GT1a, and GT1b) and neutral glycosphingolipids (Forssman antigen, globoside, and CTH) containing various fatty acids were analyzed by positive and negative ion fast atom bombardment mass spectrometry (POS-FAB-MS and NEG-FAB-MS) without any derivatization. NEG-FAB-MS of the molecules gave intense peaks of the molecular ion species, (M-H)-, and many fragment ions useful for the elucidation of carbohydrate structure were also detected with significant intensity. The fragment ions were assessed to be cleaved at the glycosidic linkages sequentially from the non-reducing end with or without the ceramide portion and it was possible to distinguish structural isomers having different binding positions of sialic acid. All molecular ion species of neutral glycosphingolipids with various fatty acids were detected at intensities that were in accordance with the relative abundance of the fatty acids.抄録全体を表示PDF形式でダウンロード (717K) -
Masakazu HIRASAWA, Goro TAMURA1984 年 95 巻 4 号 p. 983-994
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーFerredoxin-dependent glutamate synthase (native enzyme) [EC 1. 4. 7. 1] of spinach has been purified to homogeneity in the presence of 2-oxoglutarate and sodium chloride and the properties of the enzyme have been studied.
The molecular weight of the enzyme was estimated to be 140, 000 by gel filtration. Subunit analysis by SDS-gel electrophoresis yielded a single protein band whose molecular weight was about 170, 000. This purified enzyme showed a flavoprotein-like absorption spectrum having maxima at 279 and 438 nm with shoulders at 415 and 460 nm and a broad band around 360 nm. Fluorometric data indicated the presence of 2 mol of flavin per mol of the enzyme. Preliminary paper chromatography results indicated the presence of FAD and FMN in the purified enzyme. The enzyme also contained 4 mol of acid-labile sulfide and 4 g-atoms iron per mol of enzyme.
In the absence of 2-oxoglutarate and/or sodium chloride, the purified enzyme was separated by either DE-52 cellulose chromatography or gel filtration with Ultrogel AcA 34 into two molecular forms (modified enzymes) with considerable inactivation.
When reduced methyl viologen plus ferredoxin was used as the electron donor, the purified (native) enzyme showed high ferredoxin-dependent activity with a specific activity of 100 units/mg protein. Methyl viologen-dependent activity was negligible in the absence of ferredoxin.
Kinetic properties and results of ESR studies were described. The results indicate that ferredoxin-linked glutamate synthase of spinach leaves is an iron-sulfur flavoprotein.抄録全体を表示PDF形式でダウンロード (1507K) -
Yoshinori MORIYAMA, Tatsuya TAKANO, Shoji OHKUMA1984 年 95 巻 4 号 p. 995-1007
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーMembrane ghosts were prepared from purified lysosomes (tritosomes) of rat liver by hypo-osmotic treatment. Mg2+-ATP-driven acidification was observed in the membrane ghosts using acridine orange as a fluorescent probe of the transmembrane pH gradient (ΔpH). Its properties were the same as those of intact lysosomes reported previously (Ohkuma, S., Moriyama, Y., & Takano, T. (1982) Proc. Natl. Acad. Sci. U. S. 79, 2758-2762; Moriyama, Y., Takano, T., & Ohkuma, S. (1982) J. Biochem. 92, 1333-1336). The H+-pump was found to be electrogenic with use of bis (3-phenyl-5-oxoisoxasol-4-yl) pentamethine oxonol as a fluorescent membrane potential probe.
Alkaline Mg2+-ATPase activity was also identified on the membranes. It showed a pH maximum of pH 8.0-8.5, a Km value for ATP of 0.36mM and a Vmax of 0.41 units/mg protein at 30°C. Its activity was inhibited by dicyclohexylcar-bodiimide, tri-n-butyltin, azide and ADP, but not by ouabain or vanadate. It differed from mitochondrial F1F0-ATPase in sensitivities to N-ethylmaleimide, 7-chloro-4-nitrobenzo-2-oxa-1, 3-diazole, quercetin, and oligomycin.
Since this alkaline Mg2+-ATPase activity is very similar to the H+-pump activity in its requirement for divalent cations, substrate specificity and sensitivities to various chemicals, it may act as a proton translocase (H+-pump). Possible mechanisms of action of some chemicals, such as 4-acetamide-4'-isothiocyanatostilbene-2, 2'-disulfonic acid, that inhibited the H+-pump but not the alkaline Mg2+-ATPase, are discussed.抄録全体を表示PDF形式でダウンロード (1010K) -
Takuji SASAKI, Kazuto KOBAYASHI1984 年 95 巻 4 号 p. 1009-1017
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーTwo protein proteinase inhibitors, anti-trypsin and anti-chymotrypsin, were isolated from the hemolymph of silkworm larva, Bomby.t nnori, using conventional gel filtration and ion exchange chromatography techniques. They had similar physicochemical properties, in molecular weight (42, 000 for anti-trypsin and 43, 000 for anti-chymotrypsin), in amino acid composition, and in CD spectrum. Further comparison of these characteristics with human serum inhibitors, α-1-proteinase inhibitor and α-1-antichymotrypsin, suggested the resemblance of silkworm and human inhibitors. But the N-terminal sequences were not homologous to each other and antiserum against each silkworm inhibitor only formed a precipitin lines with its own antigen. These results indicated differences in minute parts of the inhibitors.抄録全体を表示PDF形式でダウンロード (1961K) -
Shio MAKINO, Ryuichi MORIYAMA, Takashi KITAHARA, Shozo KOGA1984 年 95 巻 4 号 p. 1019-1029
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーBovine band 3 in membrane-bound and soh.ubilized states was digested with chymotrypsin, trypsin, and papain. Bovine band 3 in red blood cells was fragmented by the proteases in a 5mM NaH2PO4--Na2HPO4 buffer containing 0.3M glucose, pH 8.0, but not in a 5mM NaH2PO4-Na2HPO4 buffer containing 0.15M NaCl, pH 8.0, in which human band 3 is cleaved by chymotrypsin and papain. When compared with the known data for human band 3, however, major fragments of bovine band 3 derived from intact cells, inside-out vesicles and unsealed ghosts were similar to those of human band 3, except that tryptic fragments were formed on the extracellular attack. The results suggest that bovine band 3 adopts a quite similar molecular arrangement in the membrane to in the human case. However, it was strongly suggested by molecular weight evaluation of fragments that the only detectable water-soluble 38, 000-39, 000 dalton fragment does not account for the entire hydrophilic pole of the band 3 molecule exposed in the cytoplasmic region of the membrane.
When isolated band 3 was treated with the enzymes in a 2% solution of nonaethyleneglycol n-dodecyl ether, the major product was indistinguishable on sodium dodecyl sulfate-gel from the water-soluble fragment of the cytoplasmic domain origin of band 3. This fragment lost its resistance to further enzymatic degradation when treated with dimethylmaleic anhydride, thus band 3 oligomers were converted into their monomers. The chymotryptic 38, 000 dalton water-soluble fragment obtained in nonaethyleneglycol n-dodecyl ether solution was a subfragment of a 50, 000 dalton piece which was produced in a 2% solution of deoxycholate after chymotrypsin treatment of band 3.抄録全体を表示PDF形式でダウンロード (4970K) -
Kenzo SATO, Keiichi HOSOKAWA1984 年 95 巻 4 号 p. 1031-1039
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーAdenovirus chromatin is constituted with three kinds of core proteins, VII, V, and μ, that are coded by the virus genome. Since a hexamer of VII contributes to formation of the nucleosome-like structure of the virion chromatin, we analyzed the interaction between DNA and VII in vitro, by the use of ultraviolet light-induced cross-linking and circular dichroism (CD) spectroscopy.
It was observed that DNA and VII in a plain mixture form a structure resembling viral chromatin. The DNA in the virion core or in the simply mixed complex appears to take a tight conformation by superfolding, based on the result that the ellipticity at 275 nm of DNA was reduced to approximately 3, 000°, and the wavelength of the positive peak was shifted from 275 to 285 nm. The change in CD spectrum caused by interaction of VII with DNA is similar to that of a protamine rather than that of a histone mixture. The interaction of VII with DNA is preferential, and VII is capable of associating more efficiently with double stranded DNA than with single stranded. The interaction is loosened by salt (0.3M NaCl) and tightened by magnesium ion. However, the interaction of a precursor core protein pro-VII with DNA was not as tight as that of VII and was not influenced by magnesium ion, presumably because of the existence of a hydrophobic processing sequence in the molecule.抄録全体を表示PDF形式でダウンロード (743K) -
Yuriko KANO, Shunji NATORI1984 年 95 巻 4 号 p. 1041-1046
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーThe fat body of Sarcophaga peregrina larvae was found to have activity for synthesis of sarcophagine (β-alanyl-L-tyrosine). This activity was due to a soluble enzyme (sarcophagine synthetase) that requires Mg2+ and ATP as cofactors. The enzyme activity decreased significantly after puparium formation and no sarcophagine syn-thesis was detected when fat body from white pupae was incubated in vitro with 3H-β-alanine. This apparent loss of sarcophagine synthesis was found to be partly due to peptidase, which was induced in the fat body after puparium formation. The activity of sarcophagine synthetase itself in the lysate of pupal fat body, however, was found to be significantly lower than that in the lysate of larval fat body, suggesting the presence of a developmentally regulated mechanism of sarcophagine production.抄録全体を表示PDF形式でダウンロード (1359K) -
Akira NAKAGAWA, Takahiro TSUJITA, Hiromichi OKUDA1984 年 95 巻 4 号 p. 1047-1054
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーA novel esterase was found in Pseudomonas fluorescens cells and purified to homogeneity as determined by polyacrylamide gel electrophoresis. The esterase was extracted from the cells by freeze-thawing and hypotonic treatment. Purification was achieved by ammonium sulfate precipitation, followed by successive chromatographies on DEAE-cellulose and benzylamine-agarose and then electrophoresis. The enzyme catalyzed the hydrolysis of methyl esters, such as methyl butyrate, but its hydrolyzing activity decreased with increase in the chain length of the alcohol moiety, and it did not catalyze the hydrolysis of triacylglycerols, such as triacetin. In contrast, the enzyme acted on various acyl residues in a series of methyl esters, such as dimethyl succinate, methyl metbacrylate, and dimethyl malate. The optimum pH for activity of this enzyme with methyl butyrate was 7.0-8.5. The enzyme was inhibited by phenylmethylsulfonylfluoride. Its molecular weight was estimated as 48, 000 by molecular sieve electrophoresis and gel filtration on Sephadex G-150.抄録全体を表示PDF形式でダウンロード (1138K) -
III. Isolation and Molecular Weight Analysis of the Component Chain of Fcγ Receptor of MacrophageYoshitomi AIDA, Kaoru ONOUE1984 年 95 巻 4 号 p. 1055-1065
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーSurface receptors of guinea pig peritoneal macrophages specific for the Fe region of IgG (Fcγ receptor) were isolated and identified as a surface-radioiodinated component with a molecular weight of 44, 000 that bound in an Fc-specific manner to IgG2 of guinea pig immunoglobulin immobilized in any of the following three different ways: IgG2 antibody in insoluble immune complex, IgG2 antibody bound to antigen-coupled Sepharose, and IgG2 covalently coupled to Sepharose. In order to obtain the Fcy receptor retaining the binding activity, the Fc-binding component was isolated by IgG2 affinity chromatography in which mild acidic buffer (pH 5.0-4.0) was chosen to elute the component bound to the affinity column. Forty-five to sixty-two percent of the eluted radioactivity was shown to rebind to the IgG2-affinity column. The bound fraction showed a single radioactive peak of 44, 000 daltons in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The Fc-binding component isolated by the affinity chromatography behaved similarly in gel filtration in the presence of a detergent, as did the detergent-solubilized Fcy receptor before isolation by affinity chromatography. These results suggested that the Fcy receptor was isolated in a native form. Furthermore, it was confirmed that the isolated Fcy receptor is distinct from actin or the actin-like protein (DNase I-binding protein) which had been reported to bind to IgG-affinity column.抄録全体を表示PDF形式でダウンロード (1476K) -
Yoshitomi AIDA, Kaoru ONOUE1984 年 95 巻 4 号 p. 1067-1072
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーCrosslinking of monomeric IgG2 molecules bound to the Fcγ receptors on the cell surface of guinea pig macrophages generated the triggering signal for the superoxidegenerating system. A binding experiment indicated that macrophages have saturable binding sites for monomeric IgG2. Scatchard analysis of the binding data showed that macrophages have an average of 4×105 binding sites per cell and the association constant for the binding was 4.2×106 M-1. Binding of monomeric IgG2 to macrophages could be detected by subsequent reaction with the 125I-labeled F(ab')2 fragment of rabbit antibody specific for guinea pig Fab. Although binding of IgG2 monomer to Fc receptor did not stimulate superoxide release, further addi-tion of the F(ab')2 fragment of anti-guinea pig Fab antibody did induce generation and release of superoxide, and the amount released was dependent on the dose of cell-bound IgG2. When macrophages were bound with a constant dose of IgG2 monomer in the first step, the superoxide release triggered by the addition of the F(ab')2 of anti-guinea pig Fab was dependent on the dose of the F(ab')2 fragment added. These results show that crosslinking of Fc receptors triggers the superoxide generation.抄録全体を表示PDF形式でダウンロード (433K) -
Tarou OGURUSU, Akio MAEDA, Tôru YOSHIZAWA1984 年 95 巻 4 号 p. 1073-1082
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーHalorhodopsin in the membrane fragments of Halobacterium halobium Y1 showed an absorption band at 576 nm, the intensity of which decreased on irradiation with red light at 0°C (Ogurusu, T., Maeda, A., Sasaki, N., & Yoshizawa, T. (1981) J. Biochem. 90, 1267-1273). Using this photobleachable property as the basis for an assay of halorhodopsin, we purified halorhodopsin by octyl-Sepharose column chromatography after extracting it from the membrane with Triton X-100. In NaDodSO4-polyacrylamide gel electrophoresis, hR appeared as a major band with an apparent molecular weight of 22, 000, but the preparation still showed several other faint bands. The purified halorhodopsin showed a main absorption band at 576 nm and a small band at around 415 nm in 1 M NaCl. The photoreactions of the purified halorhodopsin at 0°C and at -75°C were similar to those of halorhodopsin in membrane fragments. Irradiation of the purified halorhodopsin with red light at 0°C resulted in a decrease of absorbance at around 576 nm with a concomitant increase of absorbance at around 410 nm. A hypsochromic photoproduct was obtained on irradiation with 650 nm light at -75°C. The dependency of the absorption spectrum of halorhodopsin on the concentration of chloride indicates that halorhodopsin has a single chloride binding site, occupation of which is responsible for modifying the spectrum.抄録全体を表示PDF形式でダウンロード (3426K) -
Hiroshi MASUNO, Takeshi OHARA, Akio IDE, Hiromichi OKUDA1984 年 95 巻 4 号 p. 1083-1090
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーThe effects of various synthetic peptides on basal and ACTH-stimulated lipolysis in fat cells were examined. Glu-Arg-Gly-Phe-Phe-Phe possessed lipolytic activity and increased ACTH-stimulated lipolysis at concentrations higher than 0.5 μmol/ml. Glu-Arg-Gly-Phe-Phe-Tyr did not cause any release of FFA from fat cells. Glu-Arg-Gly-Leu-Leu-Leu had no lipolytic activity but inhibited ACTH-stimulated lipolysis at concentrations higher than 0.25 μmol/ml. Glu-Arg-Gly-Leu-Leu-Leu also inhibited epinephrine-stimulated lipolysis.
The effects of the peptides on basal and insulin-stimulated lipogenesis in fat cells were examined. Glu-Arg-Gly-Phe-Phe-Tyr increased both basal and insulinstimulated lipogenesis. A tripeptide, Glu-Arg-Gly, inhibited both basal and insulinstimulated lipogenesis. Glu-Arg-Gly-Leu-Leu-Leu had no effect on either basal or insulin-stimulated lipogenesis. Glu-Arg-Gly-Phe-Phe-Phe and ACTH, which elicit FFA release from fat cells, also stimulated formation of [14C] triglyceride from [14C] -glucose.抄録全体を表示PDF形式でダウンロード (586K) -
Katsuhiro SAGISAKA, Mamoru TAMURA, Isao YAMAZAKI1984 年 95 巻 4 号 p. 1091-1103
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリー1. From experiments at various perfusion pressures in hemoglobin-free perfused rat hearts, oxygen consumption and redox shift of pyridine nucleotide were found to vary linearly with cardiac work. This relation was used for analysis of the energy metabolism associated with ion pumps.
2. Mechanical activities such as left ventricular pressure and heart rate varied with the extracellular K+ concentration. Ion-pump dependent changes in oxygen consumption and redox state of pyridine nucleotide, estimated as the difference of the values at normal (4.7mM) and various other extracellular K+ concentrations with corrections for the change due to mechanical work, were found to vary linearly with the K+ concentration. The slope for oxygen consumption was about 0.1 μmol/min/g•wet wt per mM K+.
3. Lactate release changed markedly but transiently, about 1min after changing the extracellular K+ concentration, and its amount varied linearly with the K+ concentration. In the steady state, however, lactate release was almost independent of the extracellular K+ concentration, although oxidized pyridine nucleotide increased with increasing K+ concentration.
4. Coronary flow increased with the extracellular K+ concentration. Heart rate changed little between 1 and 12mM K+, but decreased sharply above 12mM K+. At 20mM K+, heart beat was arrested and approximately 40% of myoglobin was deoxygenated. The intracellular oxygen concentration was estimated to be about 10 μM even during aerobic perfusion. Similarly, Ca2+-free arrested heart was found to be in a hypoxic state. The results showed that oxygen entry into cardiac tissue is facilitated by the cardiac cycle.抄録全体を表示PDF形式でダウンロード (941K) -
Koei HAMANA, Shigeru MATSUZAKI1984 年 95 巻 4 号 p. 1105-1110
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーWe analyzed the cellular contents of not only major polyamines but also minor polyamines in slime molds Phvsaruni polveephalum and Dictyostelium discoideum. The presence of putrescine and spermidine in either plasmodia or myxamoebae of these molds as major polyamines was confirmed. In addition to these polyamines, appreciable amounts of 1, 3-diaminopropane were detected in P. polvicephalum and D. discoideum. Cadaverine and sym-homospermidine were detected in P. polycephalum even when the slime mold was cultured in a chemically defined growth medium. Spermine was not detected when these molds were grown in synthetic media. Other “unusual” polyamines such as norspermidine, norspermine, thermospermine, aminopropylcadaverine, and canavalmine were not detected in either mold.抄録全体を表示PDF形式でダウンロード (1033K) -
Makoto KINOSHITA, Mitsuyo OKAZAKI, Hirokazu KATO, Tamio TERAMOTO, Teru ...1984 年 95 巻 4 号 p. 1111-1118
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーA simple and rapid method for the analysis of apolipoproteins in high density lipoprotein (HDL) by high performance liquid chromatography (HPLC) has been developed (Kinoshita et al. (1983) J. Biochem. 94, 615-617). With this method, using a sodium phosphate buffer containing 0.1% sodium dodecyl sulfate (SDS) as an eluent, apolipoproteins can be analyzed from a very small amount of HDL fraction without delipidation using organic solvents. Separation profiles of apolipoproteins by this method were examined using several techniques. The elution pattern monitored by A280 can give precise quantitative as well as qualitative information about size-distribution of apolipoproteins, except for the apo C group. Moreover, separation of apo E from apo A-I was found to be improved by column elongation.抄録全体を表示PDF形式でダウンロード (1184K) -
Hisashi NAGAMOTO, Koichi YAGI1984 年 95 巻 4 号 p. 1119-1130
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーMyosin light chain kinase was prepared from rabbit skeletal muscle. DEAE-Sephadex, calmodulin-Sepharose 4 B affinity gel and Ultrogel AcA 34 were used for the purification. It took 3 days for the preparation, and 6.2mg of myosin light chain kinase was isolated from 600g of frozen muscle. The molecular weight of the myosin light chain kinase estimated by sedimentation equilibrium analysis was 103, 000±4, 100. The isoelectric point was 5.0. Chemical modification of cysteine residues did not affect the catalytic activity, but modification of tyrosine residues diminished the activity. In order to activate myosin light chain kinase, it was necessary to bind calmodulin in an equimolar ratio and the dissociation constant was estimated to be 3.6 nM. The optimum pH for the catalytic activity was 7.5, and the activity was inhibited by NaCI and KCI. In the presence of 2.74mg/ml myosin light chain and 75mM KCl, the catalytic activity was found to be 88 s-1. The Vm, and Km at 0.14M KCl were 100 s-1 and 53 μM, respectively, for the isolated light chain as substrate and 70-80 s-1 and 19 μM for myosin as substrate.抄録全体を表示PDF形式でダウンロード (923K) -
Genichiro OSHIMA1984 年 95 巻 4 号 p. 1131-1136
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーThe amidolytic activity of chymotrypsin for Suc-Ala2-Pro-Phe-MCA was somewhat enhanced by dimyristoyl PC at low ionic strength, but not at high ionic strength. The activity was strongly inhibited by pure egg yolk PA. The inhibition by 200 ng PA was neutralized by addition of 1 μg dimyristoyl PC or pure egg yolk PC, which formed vesicles with the PA.
The Km and kcat (s-1) values of chymotrypsin for hydrolysis of Suc-Ala2-Pro-Phe-MCA changed from 15 μm to 42 μM, 0.1mM and 0.5 mM, and from 1.5 to 2.7, 3.7, and 1.0 in the presence of 1 μg dimyristoyl PC, 0.5 μg pure egg yolk PE and 0.2 μg egg yolk PA, respectively.
Gel-filtration chromatography showed that dimyristoyl PC formed a complex with chymotrypsin, but did not interact with the substrate, indicating that the basic globular protein, chymotrypsin, interacted with net-neutral PL.抄録全体を表示PDF形式でダウンロード (485K) -
Kazuo NAKAMURA, Ryojiro FUJITA, Kunihiro UENO, Shizuo HANDA1984 年 95 巻 4 号 p. 1137-1144
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーNeutral and acidic glycosphingolipids were purified from porcine pancreas by chromatography on columns of DEAE-Sephadex and latrobeads. The chemical structures of the purified glycolipids were determined by carbohydrate analysis, methylation analysis, enzyme treatment, fatty acid analysis, NMR and IR. The major glycolipid of porcine pancreas was Gal (α, 1-4) Gal (β, 1-) ceramide. Gangliosides GM3 and GD3 were major acidic components and galactosylceramide 3-sulfate was also found.抄録全体を表示PDF形式でダウンロード (3595K) -
IV. Structure Required for Phospholipids for Reconstitution of the Delipidated Fc Receptor of MacrophagesMari ITONAGA, Yoshitomi AIDA, Kaoru ONOUE1984 年 95 巻 4 号 p. 1145-1153
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーTo analyze the interaction of the macrophage Fe receptor with phospholipids, we established an experimental system for delipidation of Fc receptor fraction and reconstitution of the Fc receptor activity in phospholipid vesicles. The separation of FcR from membrane phospholipids was achieved by ion exchange chromatography on DEAE-cellulose of the anionic detergent-lysate of the crude membrane fraction of guinea pig macrophages in the presence of detergent. The separation was based on the difference in charge between the complex of FcR and the anionic detergent and that of phospholipids and the detergent. The FcR fraction free of phospho-lipids showed no FcR activity as assessed in terms of its ability to inhibit the binding of labeled soluble immune complex of IgG2 antibody to macrophages, but the same fraction showed a definite activity when associated with phospholipids. This fraction was shown to contain a component of 44, 000 daltons that is susceptible to surface-labeling and binds to IgG2-Sepharose in the affinity chromatography, indicating this component to be the Fc receptor. Reconstitution experiments with this fraction showed that phosphatidylcholine is the most effective phospholipid to reconstitute the FcR activity among those tested. Phosphatidylserine, phosphatidylinositol, and sphingomyelin were ineffective, while phosphatidylethanolamine showed a moderate effect. The inactivating effect of phospholipase C treatment on the Fc receptor activity of the membrane was shown to be due to the cleavage of phospholipids in the membrane but not due to modification of the Fc receptor molecule itself.抄録全体を表示PDF形式でダウンロード (801K) -
Hirotomo YAMADA, Hisashi HAYASHI, Yasuo NATORI1984 年 95 巻 4 号 p. 1155-1160
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーA procedure for the isolation of highly purified lysosomes from normal rat liver is described. The method depends on the swelling of mitochondria when the postnuclear supernatant fraction is incubated with 1mM Ca2+. The lysosomes can then be separated from the swollen mitochondria by Percoll density gradient centrifugation. The lysosomal fraction obtained by our method was enriched more than 120-fold in terms of the marker enzymes with a yield of 25%. The electron microscopic examination and the measurement of the activities of marker enzymes for various subcellular organelles indicated that our lysosomal preparation was essentially free from contamination by other organelles.抄録全体を表示PDF形式でダウンロード (1044K) -
Nobuo SEKI, Takahiro WATANABE1984 年 95 巻 4 号 p. 1161-1167
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーConnectin was isolated from fish dorsal myofibrils by an SDS-gel filtration method and estimated to account for approximately 13% of the total myofibrillar proteins. There was no significant difference in the amount of connectin among seven fish species but rabbit skeletal myofibrils contained a slightly higher content (16%) of connectin. The high molecular weight connectins from carp and rabbit both showed a doublet band, consisting of bands 1 and 2, on SDS-polyacrylamide gel electrophoresis using a large-pore gel. However, rabbit band 1 (a component of the connectin doublet) was found to migrate more slowly than carp band 1.
During post-mortem ageing of the muscles, it was observed that the band 1 component rapidly disappeared with a concomitant increase in band 2 component and then the band 2 component was transformed slowly into faster migrating components. These results suggest that post-mortem ageing has qualitatively similar effects on the submolecular compositions of carp and rabbit connectins. However, the apparent rate of disappearance of the band I component was considerably higher in carp muscle than that in rabbit muscle.抄録全体を表示PDF形式でダウンロード (2465K) -
Makoto NISHIKATA1984 年 95 巻 4 号 p. 1169-1177
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーAn enzyme was purified from soybean seeds mainly by repeated ion-exchange chromatography using benzoyl-L-arginine p-nitroanilide (BAPA) as a substrate. The purified enzyme was homogeneous as judged by disc gel electrophoresis. The molecular weight was estimated as 59, 000 by gel filtration. The enzyme was most active toward BAPA between pH 8 and 10. The enzyme was inactive toward protein substrates but hydrolyzed synthetic substrates and oligopeptides exclusively at the carboxyl side of L-arginine and L-lysine. Kinetic studies using synthetic substrates showed that, on the basis of Vmax/Km, the enzyme preferentially hydrolyzed amide substrates over ester substrates. Benzoyl-L-arginine 4-methylcoumaryl-7-amide (Bz-Arg-MCA) was the best substrate. The enzyme was strongly inhibited by diiso-propylfluorophosphate (DFP), tosyl-L-lysine chloromethyl ketone (Tos-Lys-CH2Cl), leupeptin, and antipain. p-Chloromercuribenzoate (PCMB) was only partially inhibitory. Various protein inhibitors of trypsin such as soybean trypsin inhibitor were ineffective. From the primary specificity and susceptibility to chemicals, the enzyme can be said to be a trypsin-like serine protease. Although the physiological role of the enzyme is unclear, it seems likely that it is involved in limited hydrolysis of certain physiological peptides during processing.抄録全体を表示PDF形式でダウンロード (1313K) -
Kazue AOYAMA, Tatsuo TANAKA, Soh HIDAKA, Kiichi ISHIKAWA1984 年 95 巻 4 号 p. 1179-1186
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーThe ribonucleoprotein complex consisting of 5 S RNA and the protein L5 was prepared from the large subunit of rat liver ribosomes. The RNA in the complex was digested in situ with RNase A or RNase T1. The RNase-resistant RNA fragments bound to the protein were recovered and purified by 2 D-PAGE, and their nucleotide sequences were determined in order to elucidate the binding sites of the RNA to the protein.
The results showed that the fragments had arisen from the 5'-end region (residues 1-21), from the second hairpin loop (residues 77-102) and from the 3'-end region (residues 106-120). Harsher digestion trimmed these fragments to shorter fragments. It was concluded that the minimal interactive sequences of 5 S RNA to the protein L5 were residues 13-21, residues 85-102, and residues 106-114. A part of the first hairpin loop, residues 41-52, was also suspected to interact with the protein.
These protein-binding sites of rat liver 5 S RNA were compared with those of Escherichia coli, Halobacterium cutirubrum and yeast, and their probable conservation from eubacteria to eukaryotes is discussed.抄録全体を表示PDF形式でダウンロード (2804K) -
Noboru OISHI, Hiraku SHIMADA1984 年 95 巻 4 号 p. 1187-1192
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーDNA ligase was purified about 2, 000-fold from blastulae of sea urchin, Hemicentrotus pulcherrimus, by means of 1M KCl-extraction, phosphocellulose, DEAE-cellulose, Sepharose CL-6B, and double-stranded DNA cellulose column chromatography. The purified DNA ligase had a molecular weight of 80, 000 (determined by Sephadex G-150) and a sedimentation coefficient of 4.1 S (by glycerol gradient centrifugation). The purified enzyme required ATP and Mg2+ (or Mn2+) as cofactors for activity, and was inhibited by N-ethylmaleimide. Apparent Km values for ATP, Mg2+, and Mn2+ were 4 μM, 2.7mM, and 0.3mM, respectively.抄録全体を表示PDF形式でダウンロード (495K) -
Takashi SUZUKI, Masaya HARADA, Yasuo SUZUKI, Makoto MATSUMOTO1984 年 95 巻 4 号 p. 1193-1200
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーBovine erythrocyte sialoglycoprotein (GP-2) (1) containing lactoseries oligosaccharide chains, which showed highly specific inhibition of hemagglutination by HVJ (Hemagglutinating virus of Japan, Sendai virus), was incorporated into neuraminidase-treated chicken erythrocytes which had lost their biological responsiveness to the virus. The GP-2-incorporated erythrocytes were agglutinated and lyzed again by the virus. Incorporation of 1, 900 molecules of GP-2 per asialoerythrocyte restored fairly well the susceptibility of the cells to HVJ-mediated agglutination and hemolysis. Treatment of the erythrocytes with neuraminidase again resulted in the complete abolishment of the response to HVJ. The above observations are consistent with the view that exogenous sialoglycoprotein, GP-2, can be functionally integrated into the surface membrane of asialoerythrocytes and serve as the receptor for HVJ during the initial adsorption-fusion phase of the virus infection of the target cells.抄録全体を表示PDF形式でダウンロード (1244K) -
Sueko HAYASHI1984 年 95 巻 4 号 p. 1201-1207
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーThe mechanism of the reduction of Corynebacterium sarcosine oxidase [EC 1. 5. 3. 1] by dithiothreitol (DTT) was investigated. The reduction followed biphasic kinetics with second-order rate constants of 54M-1•S-1 and 5.4M-1•S-1 for the respective phases. When the oxidized enzyme was titrated with sarcosine under anaerobic conditions, no intermediate, such as a semiquinone or a charge-transfer complex, appeared during the reduction of the enzyme. On the other hand, on DTT titration, an intermediate with a semiquinoid character appeared, and its formation was maximum when half of the total FAD was reduced.
An oxidized semiapoenzyme, which had lost 45% of the noncovalently-bound FAD present in the native enzyme, also showed biphasic kinetics in the reduction with DTT. The second-order rate constant was found to be 38M-1•S-1 for the fast phase. An intermediate was also formed and its concentration, estimated by electron spin resonance (ESR) measurement, was found to agree with that of the noncovalently-bound FAD. In addition, the oxidized semiapoenzyme, which had lost 95% of the noncovalently-bound FAD present in the native enzyme, was reduced with DTT much more slowly than the native enzyme. In this case, the second-order rate constant was found to be 0.4M-1•S-1, and no intermediate was observed during the titration with DTT.
On the basis of these data, it is suggested that the noncovalently-bound FAD accepts electrons directly from DTT in the fast phase through the semiquinoid form, while the covalently-bound FAD accepts electrons from the reduced noncovalently-bound FAD in the slow phase without forming an intermediate.抄録全体を表示PDF形式でダウンロード (1149K) -
Tsuguo MIZUOCHI, Junko AMANO, Akira KOBATA1984 年 95 巻 4 号 p. 1209-1213
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーThe susceptibility of a variety of oligosaccharides to endo-β-N-acetylglucosaminidase D was investigated. The oligosaccharides having the structures of Manα1→6 (GlcNAcβ1→4Manα1→3)Manβ1→4GlcNAcβ1→4(±Fucα1→6)GlcNACOT, derived from complex type triantennary sugar chains, released ±Fucα1→6GlcNACOT upon incubation with the enzyme at almost the same rate as Manα1→6 (Manα1→3) Manβ1→4GlcNAcβ1→4GlcNACOT. When the reaction products were reduced with NaB3H4, and analyzed by Bio-Gel P-4 column chromatography, a new radioactive peak was detected in both cases. This new radioactive oligosaccharide was confirmed to be Manα1→6 (GlcNAcβ1→4Manα1→3) Manβ1-4GlcNACOT in the former case and Manα1→6 (Manα1→3) Manβ1→4GlcNACOT in the latter. These results indicated that endo-β-N-acetylglucosaminidase D does not require the presence of a free hydroxyl group at the C-4 position of the α-mannosyl residue of the trisaccharide glycon: Manα1→3Manβ1→4GlcNAcβ1→.抄録全体を表示PDF形式でダウンロード (335K) -
Kanji HIRAI, Kazuyoshi IKUTA, Kumiko MAOTANI, Shiro KATO1984 年 95 巻 4 号 p. 1215-1218
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーMarek's disease virus (MDV) showed only 0.3-0.6% homology in DNA sequence with herpesvirus of turkeys (HVT) at Tm-24.4°C in spite of its antigenic similarity to the latter. Southern blot hybridization under stringent conditions showed that the homology between MDV and HVT is located in the restricted portion of these viral genomes. At Tm-49.6°C, which permits the detection of homology with one base mismatch in three between the MDV and HVT DNAs, sequences with weak homology were found to be distributed over most of these viral genomes. No homology was detected between Epstein-Barr virus and either MDV or HVT DNA.抄録全体を表示PDF形式でダウンロード (1680K) -
Yasuo SUZUKI, Yoshio HIRABAYASHI, Makoto MATSUMOTO1984 年 95 巻 4 号 p. 1219-1222
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーA useful method for N-deacylation of the ceramide moiety of glycosphingolipids has been developed. Galactosylceramide, glucosylceramide, lactosylceramide, and galactosyllactosylceramide were effectively deacylated by heating with anhydrous hydrazine at 150 C for 15-25 h. The lyso-derivative as the deacylated product of the ceramide moiety of each glycosphingolipid was isolated by preparative silica gel thin layer chromatography with a 70-85% yield from the starting glycolipids. Hydrazine sulfate was an effective catalyst for the deacylation of the ceramide moiety. No dissociation of oligosaccharide moieties of the glycolipids on hydrazinolysis was confirmed by gas chromatographic analysis and N-acylation of these lysoderivatives.
The free amino groups of the lysoglycosphingolipids can be combined with various kinds of probes giving useful derivatives for biochemical and immunological studies on glycosphingolipids.抄録全体を表示PDF形式でダウンロード (1807K) -
Chiri MATSUMOTO, Yukie SUMA, Kinji TSUKADA1984 年 95 巻 4 号 p. 1223-1226
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーTreatment of rats with an ethionine plus adenine or a methionine diet leads not only to a marked increase of the α-form isozyme of S-adenosylmethionine synthetase in liver, but also to the accumulation of comparable amounts of S-adenosylethionine and S-adenosylmethionine in liver. Transplantation of ascites tumor cells into mice leads to a marked increase only of the β-form isozyme in the host liver, but the levels of S-adenosylmethionine do not significantly change in liver.抄録全体を表示PDF形式でダウンロード (287K) -
Fumio FUKAI, Akira MURAYAMA1984 年 95 巻 4 号 p. 1227-1230
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーIt was recently shown that uterine estrogen receptor (ER) is translocated from the cytoplasm into the nucleus in the form of vero-ER•E (basic ER molecule bound with estradiol), and the translocation is inhibited by the specific binding of vero-ER•E with the cytoplasmic protein factors designated as ER-binding factors (ERBFs) [“5S” ER-forming factor (“5S” ER-FF), “6S” ER-FF, “8S” ER-FF] [Murayama, A. & Fukai, F. (1983) J. Biochem. 94, 511]. It was found that the specific interaction of vero-ER•E with the ERBFs is regulated by Mg2+ at physiological concentrations. The apparent Kd values of vero-ER•E for the ERBFs [“5S” ER-FF, 2.7×10-9M; “6S” ER-FF, 6.0×10-9M; “8S” ER-FF, 3.5×10-9M] observed in the presence of 1mM Mg2+ were all increased over 10 times as compared with in the absence of Mg2+. The inhibitory effects of the ERBFs on the nuclear translocation of vero-ER•E were reduced by Mg2+.抄録全体を表示PDF形式でダウンロード (282K) -
Seiichi KAWASHIMA, Mariko OHSUMI, Masami HAYASHI, Mitsushi INOMATA, Me ...1984 年 95 巻 4 号 p. 1231-1234
発行日: 1984/07/01
公開日: 2008/11/18
ジャーナル フリーCalcium-activated neutral proteases (CANPs) with high sensitivity (μCANP) and low sensitivity (mCANP) to calcium ions were purified individually from rabbit skeletal muscle and rabbit lung and compared as to their electrophoretic properties, calcium requirements and peptide mapping of fragments produced by S. aureus V 8 protease digestion of separated subunits. All of the results suggested that there is no difference between the μCANPs as well as between the mCANPs obtained from the two tissues, with respect to the chemical and enzymatic properties. However, the contents of CANPs in these tissues were different.抄録全体を表示PDF形式でダウンロード (1606K) -
1984 年 95 巻 4 号 p. 1235
発行日: 1984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 (20K)
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