巻号一覧
108 巻, 4 号
選択された号の論文の37件中1～37を表示しています
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Hideo Yamamoto, Hiroaki Ito, Hideji Nakamura, Eijiro Hayashi, Susumu K ...1990 年 108 巻 4 号 p. 505-506
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーBinding studies of human plasma gelsolin with ATP were done by equilibrium dialysis. Analysis of the binding data showed that plasma gelsolin had one class of ATP binding site with Kd =2.8×10-7M, which saturated at an ATP/gelsolin ratio of 0.6. The bioluminescent assay for ATP with luciferin and firefly luciferase confirmed that the protein contained a nucleotide as ATP.抄録全体を表示PDF形式でダウンロード (279K) -
Shinobu Watarai, Misao Onuma, Satoshi Yamamoto, Tatsuii Yasu1990 年 108 巻 4 号 p. 507-509
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーThe effect of galactocerebroside 3'-sulfate (sulfatide) or cholesterol sulfate on syncytium formation induced by bovine immunodeficiency virus (BIV)-infected cells was investigat-ed in vitro. Sulfatide was purified from bovine brain and incorporated in liposomes which were composed of egg phosphatidylcholine (PC), cholesterol (Chol), and dipalmitoylphos-phatidic acid (DPPA). Either sulfatide- or cholesterol sulfate-containing liposomes effec-tively prevented syncytium formation induced by BIV-infected cells, but the inhibitory effect of sulfatide alone on syncytium formation was low. On the other hand, neither liposomes containing galactocerebroside nor liposomes composed of egg PC, Chol, and DPPA had any effect on syncytium formation induced by BIV-infected cells. These results suggest that liposomes containing sulfatide or cholesterol sulfate are an efficient agent to inhibit syncytium formation induced by BIV-infected cells, and that sulfate residue might play an important role in the inhibition of syncytium formation.抄録全体を表示PDF形式でダウンロード (443K) -
Takayuki Odahara, Yoshimasa Kyogoku, Hideo Akutsu1990 年 108 巻 4 号 p. 510-512
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーIn order to obtain information of the supramolecular structures of nucleic acids in vivo, the 1H-31P cross-polarization NMR technique was applied to intact marine bacterial cells. An asymmetric powder pattern spectrum of nucleic acids in the cell was observed. The major contributor to the spectrum was ribosomes. Furthermore, the powder pattern changed dramatically with the physiological conditions of the cells. The results showed that this is a promising method for the investigation of the supramolecular structures of nucleic acids in vivo.抄録全体を表示PDF形式でダウンロード (336K) -
Kiichiro Matsumura, Shinichiro Hori, Hiroko Sugiura, Teruo Shimizu1990 年 108 巻 4 号 p. 513-515
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーTwo-dimensional gel electrophoresis using 0.5% agarose gel in the 1st dimension and 2-12% gradient polyacrylamide gel in the 2nd dimension succeeded in the isoelectrical focusing of connectin, a giant myofibrillar protein of _??_3, 000 kDa. Immunoblotting with an anti-connectin monoclonal antibody, SM1-36-2, following the two-dimensional gel electropho-resis, demonstrated that connectin was an acidic protein with an estimated pI of _??_5.7.抄録全体を表示PDF形式でダウンロード (2274K) -
Yuri Miura, Hideo Utsumi, Masahiro Kashiwagi, Akira Hamada1990 年 108 巻 4 号 p. 516-518
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーThe effects of oxygen on the metabolism of lipophilic nitroxide and the membrane structure of microsomes were investigated with an ESR spectrometer equipped with a hand-made gas-permeable sampling tube. The half life of gas change in the membrane was confirmed to be 48s by the line-width of the ESR signal. The signal intensity of lipophilic nitroxide in microsomal membranes decreased with NADPH under N2 and increased under O2. The change of signal intensity was reversible, suggesting that lipophilic nitroxide can be used as an indicator of the redox state in membranes for in vivo ESR imaging.抄録全体を表示PDF形式でダウンロード (327K) -
Takao Ojima, Hiroyuki Tanaka, Kiyoyoshi Nishita1990 年 108 巻 4 号 p. 519-521
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーAkazara scallop troponin-I of Mr 52, 000 (52K) was cleaved into two fragments of 17K and 35K with cyanogen bromide. The 17K fragment, along with tropomyosin, inhibited weakly the rabbit actomyosin Mg-ATPase activity, however, the 35K fragment did not affect it at all. In the presence of Akazara scallop TnT (40K component), the 17K fragment, in turn, strongly inhibited the activity, while the 35K fragment did not. The amino acid composition and partial amino acid sequence suggested that the 17K and 35K fragments were derived from C-and N-terminal regions of the TnI, respectively, and that structural similarity to TnIs from other animals is present in the 17K region.抄録全体を表示PDF形式でダウンロード (275K) -
Yoshio Imai, Tomohide Uno, Masahiko Nakamura1990 年 108 巻 4 号 p. 522-524
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーcDNA for chimeric protein, P450(3P4), consisting of the amino-terminal 43 residues (the membrane-anchor region) of rabbit P450IIC14 and the remaining 447 residues of rabbit P450IIE1 was constructed, then cloned into expression vector pAAH5, and expressed in Saccharomyces cerevisiae AH22 cells under the control of yeast ADH1 promotor. P450(3P4) thus synthesized in the transformed yeast cells was partially purified, and its spectral and catalytic properties were examined. In the oxidized state P450(3P4) exhibited a high-spin type absorption spectrum even in the absence of a substrate. The reduced CO complex of the P450 showed a Soret absorption maximum at 452 nm. P450(3P4) catalyzed aniline p-hy-droxylation, N-nitrosodimethylamine demethylation, benzphetamine N-demethylation, and laurate and caprate (ω-1)-hydroxylation in the reconstituted system containing the P450 and NADPH-P450 reductase. These results indicate that P450(3P4) preparation obtained from the transformed yeast cells has spectral and catalytic characteristics identical with those of P450IIE1 purified from rabbit liver microsomes, confirming the substrate specificity reported of P450IIE1.抄録全体を表示PDF形式でダウンロード (1014K) -
Kei-ichi Uemura, Eiko Sugiyama, Chisato Tamai, Atsushi Hara, Tamotsu T ...1990 年 108 巻 4 号 p. 525-530
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリー1-Phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), an effective inhibitor of UDP-glucose:ceramide glucosyltransferase, caused growth inhibition of cultured rabbit skin fibroblasts in a dose-dependent manner. At 50μM both threo and erythro isomers of PDMP completely suppressed the cell growth. Major gangliosides of the fibroblasts, GM3 and GD3, were greatly reduced in amounts in the presence of threo-PDMP and accumulation of ceramides was observed. Surface labeling with galactose oxidase and [3H]NaRRA demonstrated that neutral glycosphingolipids with four or more sugars present on the surface of control cells were not detectable when the fibroblasts were grown in medium containing threo-PDMP. Metabolic labeling of cellular glycosphingolipids with [14C]-galactose showed reduced incorporation of radioactivity into gangliosides and neutral glycosphingolipids when threo-PDMP was present in the medium. In contrast, the erythro isomer of PDMP did not affect the biosynthesis of glycosphingolipids, a result suggesting that the inhibitory effect of erythro-PDMP on cell growth was due to a mechanism other than the inhibition of glucosyltransferase.抄録全体を表示PDF形式でダウンロード (4043K) -
Atsushi Irie, Hideo Kubo, Fuyuhiko Inagaki, Motonori Hoshi1990 年 108 巻 4 号 p. 531-536
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーCeramide dihexoside was obtained from the spermatozoa of the starfish, Asterias amuren-sis. Gas-liquid chromatography of the methanolysate to determine the sugar composition of the lipid demonstrated an unequal ratio of glucose and galactose, implying that two or more glycolipids are present. They were separated by thin-layer chromatography on a borate-impregnated plate into two bands. From the results of metbylation analysis and chromic acid oxidation, one was determined to be lactosylceramide, while the other was suggested to be a mixture of two diglucosylceramides: gentiobiosylceramide (Glcβ1-6Glcβ1-1Cer) and cellobiosylceramide (G1cβ1-4G1cβ1-1Cer). The molar ratio of gentiobiosyl-, cello-biosyl-, and lactosylceramide was estimated to be 0.7 : 0.3 : 1.0. Ceramide dihexosides obtained from another batch of the spermatozoa, collected at the same place in a different year, consisted almost exclusively of gentiobiosylceramide as confirmed by proton-nuclear magnetic resonance spectroscopy and fast-atom bombardment mass-spectrometry. The fatty acid compositions of these glycolipids were similar and the main acids were 14h:0, 15h:0, 16h:0, 18h:0, and 24h:1 (constituting more than 80% of the total acids). The long-chain base compositions were qualitatively similar and the major constituents were commonly d18: 2, d18:3, d19: 3, d22: 2, and t22:1. Lactosylceramide was rich in t22:1, while diglucosylceramides were rich in d22:2.抄録全体を表示PDF形式でダウンロード (1428K) -
Fuminori Umenishi, Hidetaro Yasumitsu, Yoshiyuki Ashida, Jun-iti Yamau ...1990 年 108 巻 4 号 p. 537-543
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーRous sarcoma virus-transformed rat liver cell line RSV-BRL secreted a neutral proteinase in a latent precursor form with a molecular weight (Mr) of 57, 000 (57k) as a major secreted protein. This enzyme was a calcium-dependent metallo-proteinase. The proenzyme was purified from the serum-free conditioned medium of the transformed cells by affinity chromatographies on a zinc chelate Sepharose column and a reactive red agarose column. When activated by treatment with trypsin or p-aminophenylmercuric acetate (APMA) in the presence of Ca2+, the purified enzyme effectively hydrolyzed casein, fibronectin, and laminin. Type IV collagen was hydrolyzed at 37°C but not at 30°C by the enzyme, whereas type I and type III collagens were hardly hydrolyzed even at 37°C. The treatment with trypsin or AMPA in the presence of Ca2+ converted this 57k proenzyme to an active and stable enzyme with Mr 42k. In the absence of Ca2+, however, APMA converted the proenzyme to an intermediate form with Mr 45k, while trypsin digested it to an inactive peptide with Mr 30k. These results demonstrate that calcium ion is essential for the activation, activity expression, and stabilization of this metallo-proteinase. Analysis of its partial amino acid sequence and amino acid composition showed that the 57k proenzyme was identical or closely related to the putative protein transin, a rat homologue of stromelysin.抄録全体を表示PDF形式でダウンロード (2655K) -
Rikio Yoshimura, Emi Kusunose, Noboru Yokotani, Satoru Yamamoto, Ichir ...1990 年 108 巻 4 号 p. 544-548
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーWe have previously reported the isolation of two forms of cytochrome P-450 (P-450) with ω-hydroxylase activities toward prostaglandin A (PGA) and fatty acids, designated as P-450ka-1 and P-450ka-2, from kidney cortex microsomes of rabbits treated with di(2-ethylhexyl)phthalate [Kusunose, E. et al. (1989) J. Biochem. 106, 194-196]. In the present work, we have purified and characterized two additional forms of rabbit kidney fatty acid ω-hydroxylase, designated as P-450kc and P-450kd. The purified P-450kc and P-450kd had specific contents of 13 and 16 nmol of P-450/mg of protein, with apparent molecular weights of 52, 000 and 55, 000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), respectively. Both the forms showed absorption maxima at 450 nm in the carbon monoxide-difference spectra for their reduced forms. These P-450s efficiently catalyzed the ω- and ω-1)-hydroxylation of fatty acids such as caprate, laurate, myristate, and pal-mitate, in a reconstituted system containing P-450, NADPH-P-450 reductase, and phos-phatidylcholine. Cytochrome b5 stimulated the reactions to only a slight extent. They had no detectable activity toward PGA and several xenobiotics tested. The two P-450s showed different peptide map patterns after limited proteolysis with papain or Staphylococcus aureus V8 protease. P-450kd was identical to rabbit pulmonary prostaglandin ω-hydrox-ylase (P-450p-2, P-450IVA4) in its first 20 NH2-terminal amino acid sequence (Ala-Leu-Ser-Pro- Thr-Arg-Leu-Pro-Gly-Ser-Leu-Ser-Gly-Leu-Leu-Gln-Val-Ala-Ala-Leu), although these P-450s were quite different in their other properties such as apparent molecular weights, catalytic activities, inducibility, and tissue distribution. P-450kd and P-450kc were also very similar to P-450ka-1 and P-450ka-2 in their terminal amino acid sequences. The results suggest that all these rabbit renal and pulmonary ω-hydroxylases are members of the same gene family (P-450IVA gene subfamily) in the P-450 superfamily.抄録全体を表示PDF形式でダウンロード (1503K) -
Kimimitsu Oda, Toshiyuki Fujiwara, Shigenori Ogata, Yukio Ikehara1990 年 108 巻 4 号 p. 549-553
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーAntibodies which specifically recognize rat proalbumin, but not mature serum-albumin, were raised. Propeptide (NH2-Arg-Gly-Val-Phe-Arg-Arg) with an additional cysteine residue at the carboxyl terminus was conjugated to ovalbumin through either sulfide or amino group and the conjugates were injected into rabbits. Monospecific antibodies were purified on a propeptide-coupled affinity column and further immobilized as an affinity support, allowing us to purify proalbumin in a single step from rat liver microsomes. The antibodies were also used to analyze the proteolytic processing of proalbumin in primary cultured rat hepatocytes.抄録全体を表示PDF形式でダウンロード (1683K) -
Jin Konishi, Kimitoshi Denda, Tairo Oshima, Takayoshi Wakagi, Etsuko U ...1990 年 108 巻 4 号 p. 554-559
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーImmunological cross-reactivity among three types of H+-ATPases, that is, three ar-chaebacterial ATPases, the F1-ATPase from thermophilic bacterium PS3 (TF1) and the vacuolar membrane ATPase from Saccharomyces cerevisiae, was examined by means of immunoblot analyses. The three archaebacterial ATPases were very similar in im-munological cross-reactivity, suggesting that they belong to the same family of ATPases. Cross-reaction was also observed between the ATPase from Sulfolobus acidocaldarius, one of the three archaebacteria, and TF1. S. cerevisiae vacuolar ATPase reacted with the antibodies prepared against each of the three archaebacterial ATPases, but did not react with the antibody against TF1. Electron microscopic examination revealed that the oligomeric structure of Sulfolobus ATPase was very similar to that of F1-ATPase. These results, taken together, suggest that the archaebacterial ATPases share close structural similarities with the vacuolar ATPases, and, to a lesser degree, with the F0F1-ATPases.抄録全体を表示PDF形式でダウンロード (3622K) -
Masami Kusunoki, Taka-aki Ono, Tadashi Matsushita, Hiroyuki Oyanagi, Y ...1990 年 108 巻 4 号 p. 560-567
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーHigh-resolution XANES (X-ray Absorption Near Edge Structure) spectroscopy for Mn in the S1 and S2 states of the spinach photosynthetic O2-evolving complex revealed distinct features in K-edge spectra, when a high signal-to-noise (S/N) ratio of ca. 80 with a low and constant background-to-signal (B/S) ratio of 0.15 to 0.18 was attained. Six features resolved in each S-state spectrum involve a pre-edge feature due to 1s→3d transitions, a main-edge feature possibly due to ls→4s transitions and four fine structures superimposed on the principal absorption bands due to 1s→4p* transitions. The high-quality pre-edge features were analyzed according to a parametric ligand-field theory in comparison with those of some typical authentic Mn complexes. It was deduced that i) all of the four Mn ions in the S1 -state are octabedrally coordinated and two of them constitute a di-μ-oxo bridged Mn(III, III) dimeric subunit; ii) the bridged Mn(III) ions are further bridged by a deprotonated water dimer, (HOHOH)-, and coordinated by imidazole-N and carboxylate-O- on the opposite side of the Mn atom from the di-μ-oxo bridge; iii) the other two Mn ions exist in the form of Mn(III) monomeric subunits; and iv) upon the S1→S2 transition, only the bridged Mn(III, III) is oxidized to Mn(III, IV). The distinct change in the principal absorption band shape upon the S1→S2 transition is briefly discussed to obtain the XANES evidence for a tetrameric Mn-cluster.抄録全体を表示PDF形式でダウンロード (954K) -
Masato Nagaoka, Masami Muto, Koji Nomoto, Takesi Matuzaki, Tunekazu Wa ...1990 年 108 巻 4 号 p. 568-571
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーThe isolation and analysis of the polysaccharide-peptidoglycan complexes of Lactobacillus casei YIT9018 are presented. Two polysaccharide-peptidoglycan complexes, PS-PG1 and PS-PG2, were solublized from the heat-killed cell by treatment with N-acetylmuramidase. PS-PG1 was composed of glucose, rhamnose, and small amount of galactose and gluco-samine. PS-PG2 was composed of glucose, rhamnose, galactosamine, and glucosamine. The ratio by weight of these fractions was about 1 : 8. PS-PG2 was analyzed in detail. Smith degradation and deamination of this complex yielded oligosaccharide units. The results of methylation analysis of these units and intact PS-PG2 led to the most probable structure of PS-PG2:
→(2Rhaα1→3[Glcα1→6]GlcNAcβ1→3Rhaα1→3GalNAcβ1)-n
↑2 1
(→2Rhaα1→3[Glcα1→2]Rhaα)m抄録全体を表示PDF形式でダウンロード (428K) -
Tetsuroh Ichiba, Osamu Takenaka, Tatsuya Samejima, Akira Hachimori1990 年 108 巻 4 号 p. 572-578
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーThe complete amino acid sequence of the inorganic pyrophosphatase from thermophilic bacterium PS-3 was determined by automated Edman analysis of the intact protein and of peptides derived from digests obtained with lysylendopeptidase, Staphylococcus aureus strain V8 protease, and arginylendopeptidase. The monomer peptide chain comprises 164 amino acid residues and has a calculated molecular weight of 18, 792. The sequence is identical at about 46% of the amino acid positions with that of the Escherichia coli enzymes.抄録全体を表示PDF形式でダウンロード (698K) -
Hiroyuki Yano, Keiichi Aso, Akira Tsugita1990 年 108 巻 4 号 p. 579-582
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーA novel method for vapor phase acid hydrolysis of protein suitable for quantitative analysis of tryptophan is presented. The hydrolysis is carried out in vapor of a mixture made of 7M HC1, 10% trifluoroacetic acid, and 20% thioglycolic acid in the presence of indole. Reasonably good recoveries of common amino acids, including tryptophan (above 75%), were achieved.抄録全体を表示PDF形式でダウンロード (376K) -
Isao Yumoto, Yoshihiro Fukumori, Tateo Yamanaka1990 年 108 巻 4 号 p. 583-587
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーCatalase was purified to an electrophoretically homogeneous state from the facultative alkalophilic bacterium, Bacillus YN-2000, and some of its properties were studied. Its molecular weight was 282, 000 and its molecule was composed of four identical subunits. The enzyme contained two protoheme molecules per tetramer. The enzyme showed an absorption spectrum of typical high-spin ferric heme with a peak at 406 nm in the oxidized form and peaks at 440, 559, and 592 nm in the reduced form. In contrast to the typical catalases, the enzyme was reduced with sodium dithionite, like peroxidases. The enzyme showed an appreciable peroxidase activity in addition to high catalase activity. The amino acid composition of Bacillus YN-2000 catalase was very similar to those of catalase from Neurospora crassa and peroxidase from Halobacterium halobium. The catalase content in the soluble fraction from the bacterium was higher with the cells grown at pH 10 than with the cells grown at lower pHs (pH 7-9).抄録全体を表示PDF形式でダウンロード (1188K) -
Akira Ogamo, Atsuko Metori, Hideki Uchiyama, Kinzo Nagasawa1990 年 108 巻 4 号 p. 588-592
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーThe tributylammonium salt of a porcine heparin subfraction with low affinity for antithrombin III (Mr 7, 500-18, 000; anti-clotting activity, 7 USP units/mg), having degrees of sulfate substitution at D-glucosamine and L-iduronic acid residues of GlcNS 0.786, GlcN6S 0.628, and IdoA2S 0.682 mol, was reacted with 10 or 20 mol of pyridine-sulfur trioxide per mol equiv. of available hydroxyl groups in N, N-dimethylformamide at -10°C for 1h. Both chemical and NMR spectroscopic analyses revealed that sulfation proceeded exclusively at HO-6 in n-glucosamine and HO-2 in L-iduronic acid residues, according to the amount of the sulfating reagent used (GlcNS: 0.825, 0.830; GlcN6S: 0.872, 0.928; IdoA2S: 0.687, 0.749 mol, respectively). Affinity chromatography of the sulfated products on antithrombin III-Sepharose gel indicated that the polysaccharide acquired some affinity for the protein following the sulfation, as shown by the increase in the proportion of the high-affinity heparin fraction (%) from 1.1 to 6.7. Biological examination of these products indicated that sulfation at natural positions1 along with the polysaccharide chain resulted in significant increases in all the activities (blood anti-clotting, anti-Factor IIa, and anti-Factor Xa), and in the strength of intrinsic fluorescence of antithrombin III.抄録全体を表示PDF形式でダウンロード (573K) -
Terho O. Eloranta, Alex R. Khomutov, Radii M. Khomutov, Tapani Hyv&oum ...1990 年 108 巻 4 号 p. 593-598
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーAminooxy analogues of spermidine, 1-aminooxy-3-N-[3-aminopropyl] -aminopropane (AP-APA) and N-[2-aminooxyethyl]-1, 4-diaminobutane (AOE-PU), were tested as substrates or inhibitors of the enzymes involved in methionine and polyamine metabolism. Both compounds were good competitive inhibitors and poor substrates of spermine synthase, good substrates of cytosolic polyamine acetyltransferase, inactivators of S-adenosylmethionine decarboxylase and inhibitors of ornithine decarboxylase. AP-APA and AOE-PU showed K1-values of 1.5 and 186 μM as inhibitors of purified spermine synthase, and Km-values of 1.4 and 2.1 mM as substrates of the crude hepatic polyamine acetyltransferase activity. AP-APA was more potent than AOE-PU in crude enzyme preparations. Neither drug had any significant effect at 1 mM concentration on the activities of spermidine synthase, methionine adenosyltransferase, S-adenosylhomocysteine hydrolase, and methylthioadenosine phosphorylase. The results suggest that compounds of this type are valuable tools in unraveling the physiology of polyamines.抄録全体を表示PDF形式でダウンロード (845K) -
Yasushi Yamazoe, Xue Ling, Norie Murayama, Dawei Gong, Kiyoshi Nagata, ...1990 年 108 巻 4 号 p. 599-603
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーThe effects of thyroid hormone and growth hormone on microsomal testosterone 7α-hydroxylase, P-450a, were studied to understand the interaction of these hormonemediated regulations in rats. In Western blots using anti-P-450a IgG, 1.7-fold higher content of P-450a was observed in livers of female than male adult rats, while no appreciable sex-related difference was detected in prepubertal rats and rats of 24 months of age. Treatment with n-propyl-2-thiouracil or thyroidectomy of male rats increased by 2-fold the hepatic content of P-450a, but neither regimen had a significant effect on the content in female rats. Levels of P-450a in both sexes of thyroidectomized rats were decreased by the supplementation of triiodothyronine (T3, 50 μg per kg, i.p. for 7 days) to levels similar to that observed in normal male rats. Hypophysectomy also caused an increase in microsomal P-450a content in male rats. Continuous infusion of human growth hormone, which mimicked the female secretion, further significantly increased the content in hypophysectomized rats to a level similar to that observed in normal female rats. In contrast, hepatic level of P-450a in hypophysectomized male and female rats was reduced by intermittent injection, which mimicked the male secretion. Clear suppression on the level of hepatic P-450a was also observed by the treatment of hypophysectomized rats with 5 or 50 μ/kg of T3 and of hG1I-infused hypophysectomized rat with 50 μ/kg of T3. These results suggest that the dual, stimulatory and suppressive, actions of growth hormone on liver P-450a level, which were reproduced, respectively, by the administration schedules designed to mimic female- and male-type secretions, may be responsible for the female dominant expression in livers of adult rats. The present data also indicate that T3 acts at least in part directly on the liver, but not through the hypothalamus-pituitary system, to suppress liver P-450a.抄録全体を表示PDF形式でダウンロード (1675K) -
Isamu Hirashiki, Fusahiro Ogata, Norio Yoshida, Satoru Makisumi, Akio ...1990 年 108 巻 4 号 p. 604-608
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーSeeds of Wisteria floribunda contain several kinds of cysteine proteinase inhibitor (cystatin). We purified and characterized one of these inhibitors, named WCPI-3. The molecular weight of WCPI-3 was estimated to be 17, 500 and 15, 700 by gel filtration and SDS-PAGE, respectively. The isoelectric point was 5.7. WCPI-3 formed an equimolar complex with native papain and the dissociation constant was estimated to be 6.1 nM. Complex formation between WCPI-3 and Cys25-modified papain, such as S-carboxy-methylated or S-carbamoylmethylated papain, could not be observed by gel filtration or native PAGE analysis. A peptide fragment derived from WCPI-3 digested by Achromobacter proteinase (lysyl endopeptidase) had the amino acid sequence of VVAGVNYRFVLK. The VVAG sequence in this fragment corresponds to the conserved sequence QVVAG which is considered to be one of binding regions to cysteine proteinases. The amino acid sequence of the amino-terminal portion (34 residues) of WCPI-3 was highly homologous to that of oryzacystatin from rice seeds.抄録全体を表示PDF形式でダウンロード (2381K) -
Katsuhiko Ono, Hideo Nakane1990 年 108 巻 4 号 p. 609-613
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーFour flavonoids (i.e., baicalein, quercetin, quercetagetin, and myricetin), known to be inhibitors of HIV-reverse transcriptase, have been shown to be more or less inhibitory to the activities of various cellular DNA and RNA polymerases. The degree of the inhibition varied depending on the combination of the flavonoid and the enzyme species: baicalein was moderately inhibitory to DNA polymerase γ and E. coli DNA polymerase I; quercetin was strongly inhibitory to DNA polymerase β and E. coli RNA polymerase and moderately inhibitory to DNA polymerase I; quercetagetin was a potent inhibitor for all of DNA polymerases α, β, γ, and I and RNA polymerase; myricetin was a strong inhibitor of DNA polymerases α and I and RNA polymerase. However, terminal deoxynucleotidyltransferase was virtually insensitive to inhibition by these flavonoids. The inhibition by the flavonoids was due to competition with the template•primer in the case of the DNA polymerases, whereas the inhibition was due to competition with the triphosphate substrate (GTP) in the case of RNA polymerase. The K1 values of these flavonoid inhibitors for DNA and RNA polymerases was determined.抄録全体を表示PDF形式でダウンロード (546K) -
Takashi Osumi, Sadaki Yokota, Takashi Hashimoto1990 年 108 巻 4 号 p. 614-621
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーFor the analysis of the molecular mechanism of the action of peroxisome proliferators, we attempted to establish the optimal conditions for obtaining the effects of the chemicals in vitro, employing an established cell line, Reuber rat hepatoma H4IIEC3. Histochemical analyses revealed a marked increase in the number, size, and catalase content of peroxisomes in the cells cultured on a medium containing 0.5 mM ciprofibrate, a peroxisome proliferator. The activity of acyl-CoA oxidase, the initial enzyme of the peroxisomal β-oxidation system, was increased by more than 10-fold by the same treatment. Catalase was also induced significantly, whereas the activities of glutamate dehydrogenase and lactate dehydrogenase, mitochondrial and cytosolic marker enzymes, did not change upon the treatment. Immunoblotting and RNA-blotting analyses confirmed the increases in the amount of protein and mRNA for all the three enzymes of the peroxisomal β-oxidation system. Cell fractionation experiments gave a partial separation of peroxisomes from other organelles for the induced culture. Thus, H4IIEC3 cells offer a good in vitro model system of the induction of peroxisomes and peroxisomal β-oxidation enzymes by peroxisome proliferators.抄録全体を表示PDF形式でダウンロード (4034K) -
Chiseko Noda, Miho Ohguri, Koichi Matsuda, Toshikazu Nakamura, Akira H ...1990 年 108 巻 4 号 p. 622-628
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーFor study of the mechanisms regulating the induction of serine dehydratase by various hormones in rat liver [Noda et al. (1988) J. Biol. Chem. 263, 14764-14768], we cloned the gene for this enzyme from a rat genomic library. The gene spans about 7.5 kilobases and consists of nine exons and eight introns. The exon-intron boundaries are consistent with the “GT-AG” rule. Southern blot analysis of rat genomic DNA suggested the presence of one copy of serine dehydratase gene per haploid genome. The 5' end of serine dehydratase mRNA is located 148 nucleotides upstream of the initiator methionine codon, ATG, determined by primer extension analysis and S1 nuclease mapping, although an alternative transcription initiation site(s) may be located a few bases downstream. The 5' flanking region of the gene lacks typical TATA and CCAAT sequences, but contains AATAAA and CATT sequences, at -25 to -20 and -54 to -51, respectively. Furthermore, there are five GC box-related sequences. There are three putative glucocorticoid-responsive elements and two copies of the CGTCA motif of the cAMP-responsive element upstream of the promoters. The 5' flanking sequence shows more than 98% homology with that reported by Ogawa et al. [(1988) Proc. Natl. Acad. Sci. U. S. A. 85, 5809-5813], but the first exons have different sequences. Another difference is that a segment of 108 nucleotides is located just upstream of exon 5 in the sequence reported here, but is included as an exon in the sequence of Ogawa et al. The possibility to producing two species of serine dehydratase mRNAs from a single gene by transcription from different sites and alternative splicing is discussed.抄録全体を表示PDF形式でダウンロード (2531K) -
Tarou Ogurusu, Takesatoru Fukuda, Ken-Ichi Furukawa, Munekazu Shigekaw ...1990 年 108 巻 4 号 p. 629-634
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーWe examined the effect of protein kinase C (PKC)-dependent phosphorylation on Ca2+ uptake and ATP hydrolysis by microsomal as well as purified sarcolemmal Ca2+-ATPasepreparations isolated from bovine aortic smooth muscle. The phosphorylation was per-formed by treating these preparations with PKC and saturating concentrations of ATP (or ATP- γS), Ca2+, and 12- 0-tetradecanoylphorbol-13-acetate (TPA) at 37°C for 10min. In microsomes, treatment with PKC enhanced a portion of the Ca2+ uptake activity inhibitable by 10pM vanadate, by up to about 30%. On the other hand, Ca2+-dependent ATPase activity in the purified Ca2+-ATPase preparation was stimulated by up to twofold. Up to twofold stimulation by PKC was also observed for the Ca2+ uptake by proteoliposomes reconstitut-ed from purified sarcolemmal Ca2+-ATPase and phospholipids. Since these effects were evident only at Ca2+ concentrations between 0.1 to 1.0μM, we concluded that it was the affinity of the Ca2+-ATPase for Ca2+ that was increased by the PKC treatment. Under conditions in which PKC increased Ca2+ pump activity, the sarcolemmal Ca2+-ATPase was phosphorylated to a level of about 1mol per mol of the enzyme. There was good parallelism between the ATPase phosphorylation and the extent of enzyme activation. These results strongly suggest that the activity of the sarcolemmal Ca2+ pump in vascular smooth muscle is regulated through its direct phosphorylation by PKC.抄録全体を表示PDF形式でダウンロード (1567K) -
Haruo Suzuki, Etsuko B. Mukouyama, Takayuki Kamei1990 年 108 巻 4 号 p. 635-641
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーThe activity of eukaryotic initiation factor eIF-2 as to the formation of the ternary complex, eIF-2 GTP Met-tRNAf, is inhibited by N-ethylmaleimide. Our preparation of pig livereIF-2 contained α and γ subunits and was inhibited by more than 90% by N-ethylmale-imide. Using our eIF-2, we determined the sequences around the N-ethylmaleimide-reactive sulfhydryl groups, studied the effect of GDP on thesulfhydryl modification and that of NEM on the [3H] GDP binding, and examined the protective effect of GTP against the inhibition of ternary complex formation by N-ethylmaleimide. Both subunits of native eIF-2 contained [14C] N-ethylmaleimide-reactive sulfhydryl groups. One N-ethylmale-imide-reactive sulfhydryl group was in the a subunit and 4 were in the γ subunit. The sequence of the peptide of the α subunit was determined to be: Ala-Gly-Leu-Asn-Cys-Ser-Thr-Glu-Thr-Met-Pro-Ile. Two of the four [14C] N-ethylmaleimide-reactive sulfhydryl groups in the γ subunit were highly reactive, their sequences being: Ile-Val-Leu-Thr- Asn-Pro-Val-Cys-Thr-Glu-Val-Gly-Glu-Lys (γ1); Ser-Cys-Gly-Ser-Ser-Thr-Pro-Asp-Glu-Phe-Pro-Thr-Asp-Ile-Pro-Gly-Thr-Lys (γ3a). Peptide γ3a contained the consensus sequence element (AspXaaXaaGly) of GTP-binding proteins. With preincubation of eIF-2 with GDP, the incorporation of [14C] N-ethylmaleimide into the γ subunit was reduced to 40% of the control level, but the 14C-incorporation into the α subunit did not change. Moreover, [3H] - GDP-binding to eIF-2 was reduced to 43% of the control level with pretreatment of eIF-2 with N-ethylmaleimide and the pretreatment of eIF-2 with GTP prevented the inhibition of ternary complex formation by N-ethylmaleimide. These data indicate that the inhibition of ternary complex by N-ethylmaleimide is at least in part due to a decrease in GTP binding to eIF-2 on the modification of sulfhydryl groups in the γ subunit.抄録全体を表示PDF形式でダウンロード (2820K) -
Carmen Valcarce, Javier Vitorica, Jorgina Satrústegui, Jos&eacu ...1990 年 108 巻 4 号 p. 642-645
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーTitration of mitochondrial respiration against the membrane potential with the inhibitor malonate has been carried out during the perinatal period in isolated rat liver mitochon-dria. Neonatal and adult mitochondria exhibited the characteristic “nonohmic” behaviorfor the proton conductance (CmH+). In contrast, fetal mitochondria exhibited an “anomalous” “ohmic” behavior for CmH+. The calculated passive proton permeability of the membrane undergoes a profound reduction during the first postnatal hour. The results reported demonstrate that the hypothesis [Pollak, J.K. & Sutton, R. (1980) Trends Biochem. Sci. 5, 23-27] of the existence of a “leaky” mitochondria in the fetal rat liver, and of its sudden neonatal change towards a state of higher energy conservation of the proton electrochemical gradient, is correct.抄録全体を表示PDF形式でダウンロード (548K) -
Kunio Miki, Toshihide Ezoe, Akihiko Masui, Takuma Yoshisaka, Mamoru Mi ...1990 年 108 巻 4 号 p. 646-649
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーC-Phycocyanin from a red alga, Porphyra tenera, has been crystallized by the vapordiffusion procedure. Both orthorhombic and hexagonal forms were obtained from ammonium sulfate solutions, whereas only the orthorhombic form was selectively grown from sodium citrate solutions. The orthorhombic crystals are more suitable for further crystallographic work; their space group is P212121, with unit-cell dimensions of a=105, b=121, and c=184Â. The asymmetric unit comprises two (αβ)3 trimer molecules of C-phycocyanin. These crystals diffract X-rays up to about 3Â resolution.抄録全体を表示PDF形式でダウンロード (2067K) -
Ryoko Kuruto, Ryushi Nozawa, Keiichi Takeishi, Kyoko Arai, Takeshi Yok ...1990 年 108 巻 4 号 p. 650-653
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーA differentiation antigen induced in 1, 25-dihydroxyvitamin D3 (VD3)-treated HL-60 cells was identified as being comprised of the myeloid calcium binding proteins CaBP-p8 and -p14 by determining its amino acid and DNA sequence. Northern blot analysis using a DNA fragment of the gene encoding p14 as a probe indicated that the gene was not expressed in undifferentiated HL-60 cells but transcribed starting on day 1 after VD3 treatment. The level of p 14 mRNA reached a peak on day 2, then declined, and little mRNA remained on day 10, indicating that the p14 gene is activated once and then inactivated during HL-60 differentiation due to VD3. In contrast, thymidylate synthase (TSase) mRNA was present in undifferentiated HL-60 cells but disappeared quickly after VD3 treatment. Both p8 and p14 of CaBP were found at elevated levels in sera of some patients with connective tissue diseases [highly elevated in rheumatoid arthritis (RA), Sjogren's syndrome (SjS), systemic lupus erythematosus (SLE), and progressive systemic sclerosis (PSS), and moderately in polymyositis or dermatomyositis (PM/DM) and mixed connective tissue disease (MCTD)] . These results were in sharp contrast with the finding that p14 is always at a highly elevated level but little p8 is present in the sera of cystic fibrosis (CF) patients [Bruggen et al. (1988) Nature 331, 570).抄録全体を表示PDF形式でダウンロード (2217K) -
Shouichi Higashi, Shun-ichiro Kawabata, Hitoshi Nishimura, Hideyuki Fu ...1990 年 108 巻 4 号 p. 654-662
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーA murine monoclonal antibody (designated VII-M31) directed against bovine factor VII was prepared and characterized. Antibody VII-M31 inhibited the activations of both factors IX and X catalyzed by factor VIIa in the presence of tissue factor, phospholipids, and Ca2+. It possessed a strong affinity for factor VII in the presence of 5 mM Ca2+ (Kd=1.12 X 10-10 M). The immunoblotting test of other bovine proteins with the antibody, such as prothrombin, factor X, factor IX, protein C, protein S, and protein Z, in addition to human factor VII, revealed that it recognizes only a Ca2+-dependent epitope in bovine factor VII. Furthermore, this antibody VII-M31 covalently coupled with Affi-Gel allowed a simple and rapid purification of bovine factor VII. To localize the antigenic site in factor VII, various segments including a γ-carboxyglutamic acid (Gla)-domainless protein, a Gla-domain peptide and the fragments isolated from the lysyl endopeptidase digest, were prepared. Among them, the isolated Gla-domain peptide and Gla-domainless factor VII were no longer recognized by antibody VII-M31, indicating that the sequence around the cleavage site by α-chymotrypsin is required for the interaction between the antibody and factor VII. In accordance with this result, the antibody bound specifically to a Gla-containing peptide corresponding to the NH2-terminal 23-50 residues of factor VII, which contains the chymotryptic cleavage site. These results suggest that the specific epitope of this antibody is localized in the carboxy-terminal 28 residues of the Gla-domain constituting the amino-terminal portion of bovine factor VII.抄録全体を表示PDF形式でダウンロード (3689K) -
Kazuo Inaba, Michio Ono, Hideo Mohri1990 年 108 巻 4 号 p. 663-668
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーConformational changes of the β chain of the outer-arm dynein from sea urchin sperm flagella in relation to ATP hydrolysis was examined by tryptic digestion. Tryptic digestion of the β chain in the presence of 2 mM ATP (ADP) and 100μM vanadate (V1) or in the presence of 4 mM ATPγS produced different polypeptides from in the case of no addition. The difference was similar to the result previously reported for 21S outer-arm dynein heavy chains [Inaba, K. & Mohri, H. (1989) J. Biol. Chem. 264, 8384-8388]. Unlike the tryptic digestion pattern of 21S dynein heavy chains, however, the 135-kDa polypeptide was consistently produced from the β chain, even in the presence of ATP (ADP) and V1. The tryptic digestion pattern of the 21S particle reconstituted from the separated a chain, the β/ IC1 complex and the IC2/IC3 complex [Tang, W.-J.Y., Bell, C.W., Sale, W.S., & Gibbons, I.R. (1982) J. Biol. Chem. 257, 508-515] was similar to that of intact 21S dynein; the 135-kDa polypeptide was only slightly produced in the presence of ATP and V1. The digestion rate constant of the 135-kDa polypeptide from the β chain in the presence of ATP and V1 was significantly decreased as compared with in the case of 21S dynein or that of the reconstitut-ed 21S particle. These results suggest that the trypsin sensitivity of the 135-kDa region of the β chain changes with the association of the β/IC1 complex with the β chain and the IC2/ IC3 complex in the presence of ATP and V1.抄録全体を表示PDF形式でダウンロード (3976K) -
Misao Tashiro, Toshio Asao, Chikage Hirata, Kyoko Takahashi, Masao Kan ...1990 年 108 巻 4 号 p. 669-672
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーThe complete amino acid sequence of a major trypsin inhibitor (FMTI-II) from seeds of foxtail millet (Setaria italica) was determined by analysis of peptides derived from the reduced and S-carboxymethylated protein by digestion with TPCK-trypsin and Sta-phylococcus aureus V8 protease. FMTI-II consists of 67 amino acid residues, including 10 half-cystine residues which are involved in 5 disulfide bridges in the molecule. The established sequence had a high degree of homology to Bowman-Birk type inhibitors from leguminous and gramineous plants. The trypsin reactive-site peptide bond in FMTI-II also appears to be Lys (16)-Ser (17) by comparison with these sequences.抄録全体を表示PDF形式でダウンロード (364K) -
Kiyoshi Yasukawa, Takashi Saito, Takeshi Fukunaga, Yasuo Sekimori, Yas ...1990 年 108 巻 4 号 p. 673-676
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーAn immunosorbent assay system to detect genetically engineered IL-6 receptor (IL-6R) was established, whereby soluble IL-6 receptor (sIL-6R) was detected in the culture medium when sIL-6R cDNA was transfected into COS1 cells. A stably transformed Chinese hamster ovary (CHO) cell line constitutively expressing sIL-6R has been established. The recom-binant sIL-6R was purified to homogeneity by sequential filtration and chromatography of the culture medium. The recombinant sIL-6R augmented the sensitivity of M1 cells to IL-6 in growth inhibition assay in a dose-dependent manner. Furthermore, a radioisotope immunosorbent assay (RIA) utilizing recombinant sIL-6R was established which could detect IL-6 in a quantity as low as low as 10 ng/ml.抄録全体を表示PDF形式でダウンロード (951K) -
Kiyoyoshi Nishita, Takao Ojima1990 年 108 巻 4 号 p. 677-683
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーTroponin was isolated from the abdominal muscle of the American lobster (Homarus americanus) by essentially the same method as used for akazara scallop troponin [J. Biol. Chem., 261, 16749-16754 (1986)]. The thus isolated troponin together with lobster tropomyosin confers high Ca2+ - sensitivity to rabbit reconstituted actomyosin. The troponin consists of components having Mr of about 42, 000, 32, 000, 30, 000, and 17, 000, but not the Mr 52, 000-59, 000 component previously reported to be present in several crustacean troponins. These troponin components were separated from each other by DEAE-Toyopearl column chromatography in the presence of 6M urea. The Mr 17, 000 component was further separated into one major and two minor components by the same chromatography, but each of them was confirmed to be a Ca2+ binding component, TnC. The Mr 32, 000 and 30, 000 components were both regarded as inhibitory subunits, TnIs, since the Mg-ATPase activity of actomyosin in the presence of tropomyosin was strongly inhibited by the addition of the components, and the inhibition was reversed by the further addition of TnC. Finally, the Mr 42, 000 component was regarded as TnT, since this component formed stoichiometric complex with TnC and TnI, and was indispensable for Ca2+ regulation of the actomyosin-tropomyosin system.抄録全体を表示PDF形式でダウンロード (3379K) -
Keiko Nohara, Minoru Suzuki, Fuyuhiko Inagaki, Hiroyasu Ito, Kunimitsu ...1990 年 108 巻 4 号 p. 684-688
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーA unique fucoganglioside was isolated from rat spleen and characterized by compositional analysis, methylation analysis, exoglycosidase treatment, negative ion fast atom bombardment (FAB) mass spectrometry, and proton NMR spectrometry. The ganglioside was identified as αGal, Fuc-GM1(NeuGc), which has the blood group B determinant at the nonreducing termini, as shown below:
This is the first report describing the occurrence in nature of αGal, Fuc-GMI containing N-glycolyIneuraminic acid.抄録全体を表示PDF形式でダウンロード (1353K) -
Yoshinori Moriyama, Masatomo Maeda, Masamitsu Futai1990 年 108 巻 4 号 p. 689-693
発行日: 1990/10/01
公開日: 2008/11/18
ジャーナル フリーEnergy coupling of L-glutamate transport in brain synaptic vesicles has been studied. ATP-dependent acidification of the bovine brain synaptic vesicles was shown to require Cl-, to be accelerated by valinomycin and to be abolished by ammonium sulfate, nigericin or CCCP plus valinomycin, and K+. On the other hand, ATP-driven formation of a membrane potential (positive inside) was found to be stimulated by ammonium sulfate, not to be affected by nigericin and to be abolished by CCCP plus valinomycin and K+. Like formation of a membrane potential, ATP-dependent L- [3H] glutamate uptake into vesicles was stimulated by ammonium sulfate, not affected by nigericin and abolished by CCCP plus valinomycin and K+. The L- [3H] glutamate uptake differed in specificity from the transport system in synaptic plasma membranes. Both ATP-dependent H+ pump activity and L-glutamate uptake were inhibited by bafilomycin and cold treatment (common properties of vacuolar H+-ATPase). ATP-dependent acidification in the presence of L-glutamate was also observed, suggesting that L-glutamate uptake lowered the membrane potential to drive further entry of H+. These results were consistent with the notion that the vacuolar H+-ATPase of synaptic vesicles formed a membrane potential to drive L-glutamate uptake. ATPase activity of the vesicles was not affected by the addition of Cl-, glutamate or nigericin, indicating that an electrochemical H+ gradient had no effect on the ATPase activity.抄録全体を表示PDF形式でダウンロード (623K)
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