巻号一覧
105 巻, 5 号
選択された号の論文の34件中1～34を表示しています
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Shigehiro Kamitori, Yoshihiko Odagaki, Katsura Inoue, Seiki Kuramitsu, ...1989 年 105 巻 5 号 p. 671-672
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーThe branched-chain amino acid aminotransferase of Escherichia coli was crystallized in two crystal systems, monoclinic and tetragonal, from polyethylene glycol and ammonium sulfate solutions, pH 7.0, respectively. The crystals were of good quality, with diffractions extending beyond 2.8 Å. The space group and unit cell dimensions of the monoclinic system crystals were determined from precession photographs to be C2, and a=93.9, b=143.6, c=143.9Å and, β=134.3°. For the tetragonal system crystals, the possible space group P422 or P4122, and cell dimensions of a=b=101 Å and c=24Å were determined. Three identical subunits exist per an asymmetric unit in both types of crystals.抄録全体を表示PDF形式でダウンロード (930K) -
Harutoshi Kizaki, Hideo Shimada, Yuzuru Ishimura1989 年 105 巻 5 号 p. 673-675
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーA phorbol ester, 12-O-tetradecanoylphorbol 13-acetate (TPA), induced the cleavage of nuclear DNA at linker regions in cultured mouse thymocytes. Similar DNA fragmentation was induced by 1-oleoyl-2-acetyl-glycerol, a synthetic diacylglycerol, but not by 4α-phorbol-12, 13 didecanoate. The DNA fragmentation was inhibited by 1-(5-isoquinoline-sulfonyl)-2-methyl-piperazine dihydrochloride, an inhibitor of protein kinase C, as well as actinomycin D and cycloheximide. It appears that TPA induces DNA cleavage through activation of protein kinase C and synthesis of yet unidentified protein (s). That the inhibition of DNA fragmentation was accompanied by a reduction in cell lysis suggests a causal relationship between DNA fragmentation and cell death.抄録全体を表示PDF形式でダウンロード (1435K) -
Toshihide Hashimoto, Ken-ichirou Morohashi, Tsuneo Omura1989 年 105 巻 5 号 p. 676-679
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーWe isolated 4 different clones of the P-450(11β) gene from a bovine genomic library. These genomic clones were highly homologous with each other. Two of the isolated clones were pseudogenes. Determination of its nucleotide sequences indicated that the bovine P-450(11β) gene is divided into 9 exons by 8 introns and that it is about 8.5 kb in total length. The number of exons and the locations of intron insertion into the P-450(11β) gene are identical with those in the case of P-450(SCC), but different from those of other microsomal P-450s.抄録全体を表示PDF形式でダウンロード (1529K) -
Michiko Sekine, Mitsuru Sakaizumi, Kazuo Moriwaki, Tamio Yamakawa, Ake ...1989 年 105 巻 5 号 p. 680-683
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーThe gene (Gsl-5) controlling the expression of GL-Y (Ga1β1-4(Fucαl-3)G1cNAcβl-6(Galβ1-3)Gb4Cer) in mouse kidney was suggested to belocated near Eα-4 on mouse chromosome 19 by the results of glycolipid analysis of BXD/Ty recombinant inbred strains (Sekine et al. [1987] J. Biochem. 101, 563-568). In this study, Gsl-5 was mapped on mouse chromosome 19. Among 133 backcross progeny produced on mating between DBA/2 mice and (WHT/Ht×DBA/2)F1 mice, 10 recombinants between Lyt-1 and Gsl-5 were detected, indicating that Gsl-5 is located at 7.5±2.3 centimorgans (cM) from Lyt-1. While among 154 backcross progeny produced on mating between DBA/2 and (DBA/2×Mus musculus castaneus)F1 mice, 39 recombinants between Got-1 and Gsl-5 were obtained, indicating that the distance between Got-1 and Gsl-5 is 25.3±3.5cM and that Gsl-5 is telomeric to Lyt-1. In the latter mating experiment, we detected 3 recombinants between Gsl-5 and the gene (Gsl-6) controlling the expression of the Z1 ganglioside (NeuGcα2-3Galβl -3Gb4Cer) among the 154 backcross mice. These results indicate that these two genes, Gsl-5 and Gsl-6, are closely linked to each other, being 1.9±1.1cM apart. This is the report of evidence that two genes controlling the expression of carbohydrates in glycoconjugates are closely linked and the first to suggest that some genes controlling the expression of carbohydrates may be clustered. All the results together showed that the genes are located in the order of Lyt-1-Gs1-6- Gs1-5- Ea-4 on mouse chromosome 19.抄録全体を表示PDF形式でダウンロード (1682K) -
Sumiko Hamanaka, Chidori Asagami, Minoru Suzuki, Fuyuhiko Inagaki, Ake ...1989 年 105 巻 5 号 p. 684-690
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーHuman epidermis gave two glycolipid bands that migrated faster than glucosylceramide and two bands that migrated like glucosylceramide and galactosylceramide, respectively, on TLC. The two faster migrating glycolipids (GL-I and GL-II.), which exhibited alkali-lability, were purified by conventional DEAE and silica gel column chromatographies, and further by HPLC on a silica gel column. Structure determination of the two components, named GL-13 and GL-II3, which were finally purified from GL-I and GL-II, respectively, by HPLC on a reversed phase column, was performed by means of 1H-NMR spectroscopy, fast atom bombardment mass spectrometry, and component analysis involving GLC-mass spectrometry. GL-13 was determined to be a mixture of glucosyl β1-N-(ω-0-linoleoy1)- triacontanoyl- and -dotriacontamonoenoyl-eicosasphingenine, and one of the two compo-nents of GL-II3 was determined to be glucosy1 β1-N-(ω-O-linoleoyl)-ptriacontanoyl-tri-hydroxyeicosasphingenine. GL-13 and GL-II3 were the major components of GL-I and GL-II, respectively, and both the latter contained additional four components, which were heterogeneous as to the ceramide portion. This paper reports the structures of acylglucosyl-ceramides isolated from human epidermis together with 1H-NMR spectra and mass spectra demonstrating their molecular weights. The structure of molecular species containing trihydroxysphingosine having a double bond is novel.抄録全体を表示PDF形式でダウンロード (2717K) -
Seiko Shigeta, Kazuhisa Ono, Masaya Ohta, Yuji Kobayashi, Satoru Oka1989 年 105 巻 5 号 p. 691-696
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーUsing the methods of methylation/GLC, MS, and 1H-NMR analyses, a disaccharide struc-ture of GalNAcα1→2Fucl→ was identified as one of the major structural units at non-reducing terminals of a highly branched and N-glycosidically linked carbohydrate on an allergenically active glycopeptide, Gp-2 (Mr ca. 8, 000). Gp-2 was one of the major fractions in the Pronase digest of a sea-squirt antigen, Gi-rep, and is capable of eliciting the skin reaction specific to patients with sea-squirt allergy similarly to Gi-rep. An additional disaccharide structure of GaINAc1→(3/4)HexNAc1→ was also expected as the other major non-reducing terminal unit, though the anomeric configuration of the terminal Ga1NAc could not be identified. As Gp-2 carbohydrate was found to contain 3 mol each of the two types of terminal Ga1NAc-containing units, both were nominated as possible components of the IO4- oxidation-sensitive epitopes which are responsible for the allergenic activity in Gp-2 and its mother antigen, Gi-rep. A few moles of Fuc and Gal were also detected as minor non-reducing terminals in addition to the 6 mol of the Ga1NAc-containing units.抄録全体を表示PDF形式でダウンロード (723K) -
Fuyuki Iwasa, Mayumi Furuya, Yujiro Hayashi, Norio Ohnuma1989 年 105 巻 5 号 p. 697-699
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーThe effects of α-rat atrial natriuretic peptide (α-rANP) and sodium nitroprusside on the activity of rat lung particulate guanylate cyclase were examined. The particulate guanylate cyclase in partially purified rat lung membranes was stimulated by both α-rANP and nitroprusside. The effects of α-rANP and nitroprusside were, however, not additive. Diamide and N-ethylmaleimide almost completely abolished the nitroprusside-mediated stimulation, while they had only moderate effects on the α-rANP-mediated stimulation of the enzyme activity. ATP potentiated the enzyme stimulation by α-rANP, whereas it had no effect on the nitroprusside-mediated stimulation. These findings suggest that the stimula-tion of lung particulate guanylate cyclase activity by α-rANP and nitroprusside is mediated by different mechanisms.抄録全体を表示PDF形式でダウンロード (1687K) -
Hiroyasu Nakata1989 年 105 巻 5 号 p. 700-704
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーA 5'-N-ethylcarboxamido[3H] adenosine ([3H]NECA) binding site of mouse mastocytoma P815 cell membranes has been purified _??_100-fold by affinity chromatography. This adenosine binding site, which has a similar specificity to that of the A2 adenosine receptor, was adsorbed on NECA-linked Sepharose 6B and eluted with NECA. The adsorption of the [3H] NECA binding site to the affinity matrix was specifically blocked by NECA. The [3H]-NECA binding site bound on the affinity matrix was also specifically eluted by NECA. This affinity matrix adsorbed _??_90% of the digitonin-solubilized [3H] NECA binding activity applied, and after the gel was washed, 30-50% of the adsorbed binding activity could be eluted with 500μM NECA with specific binding activity of 50-70 pmol/mg of protein. The affinity-purified [3H] NECA binding site retained the same ligand binding specificities as the original membrane preparation. The results indicate that the NECA-Sepharose 6B should provide a powerful tool for the eventual purification of [3H] NECA binding sites of P815 cell membranes.抄録全体を表示PDF形式でダウンロード (641K) -
Snezana R. Vrbaški, Vanja I. Ristic, Gordana T. Petrovic, Milos ...1989 年 105 巻 5 号 p. 705-707
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーMale Wistar rats with an initial weight of 170g were maintained on a nutritionally adequate diet, and diazepam was administered at a dose of 10 mg/kg/d. Control animals were pair-fed an adequate diet. The feeding was continued for 180 d, and the effects on brain lipid contents were studied. It was found that the contents of the phospholipids, phos-phatidylethanolamine and phosphatidylserine, the monogalactosyl glycolipids, hydroxy and nonhydroxy fatty acyl galactocerebroside, sulfoglycolipids, and the gangliosides, GM1, GD1a, GD1b, and GT1b, were significantly reduced in the brain of diazepam-treated rats. There was a significantly increased content of phosphatidylinositol after 180 d of diazepam treatment. The results suggest that changes in brain lipid content may mediate the adaptive changes that occur upon prolonged exposure to diazepam.抄録全体を表示PDF形式でダウンロード (394K) -
Nobuyuki Yamasaki, Nural Absax, Gunki Funatsu1989 年 105 巻 5 号 p. 708-711
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーThe binding of saccharides to Abrus precatorius agglutinin (APA) was analyzed by fluorescence spectroscopy. Upon binding of specific saccharides, the fluorescence emission maximum of APA (338nm) shifted to shorter wavelength by 5 nm, owing to the change in the environment of tryptophan. By analyzing the change in the fluorescence intensity at 338 nm as a function of concentration of saccharides, the association constants for binding of saccharides to APA were determined. The results suggest that in the saccharide binding site on each B-chain of APA, there may be a site which interacts with the saccharide residue linked to galactopyranoside at the non-reducing end, in addition to the site which recog-nizes the galactopyranosyl residue. Fluorescence quenching data indicate that 8 out of 24 tryptophans in APA are located at or near the surface of the protein molecule and are available for quenching with both KI and acrylamide, and 10 tryptophans are involved in the environment to which acrylamide has access but KI does not. Binding of lactose to APA reduced by 4 the number of tryptophan residues accessible to quenchers. Based on the results, it is suggested that the tryptophan residues at the saccharide binding site on each B-chain of APA are present on the surface of the APA molecule, and they are shielded from quenching by KI and acrylamide upon binding with specific saccharides.抄録全体を表示PDF形式でダウンロード (527K) -
Masaki Hosoya, Jun-Ichi Miyazaki, Tamio Hirabayashi1989 年 105 巻 5 号 p. 712-717
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーIn the embryonic smooth muscle of chicken gizzards we found 4 high-Mr-type and 5 low-Mr-type tropomyosin isoforms in addition to α-and β-isoforms reported already. The criteria by which they were identified as tropomyosin isoforms were as follows: 1) anomalous reduction of electrophoretic mobility in the presence of urea, 2) cross reactivity with antisera against tropomyosins, 3) inclusion in a tropomyosin preparation obtained by the usual method for tropomyosin purification, and 4) binding ability to skeletal muscle actin. At the early stages of development, the amounts of these isoforms were larger than those of α-and β-isoforms, but they gradually decreased at later stages and finally disappeared completely after hatching. Our previous study of gizzard smooth muscle showed that the amount ratio of accumulated tropomyosin to γ-actin was reasonably constant in the development after hatching, while, at the earlier embryonic stages (7-14 d of incubation), it was lower than expected. The isoforms found in this study were present in amounts large enough to bring the ratio at the earlier stages up to the constant amount ratio observed after hatching. Therefore, the coordinate accumulation of actin and tropomyosin was suggested to occur even at the embryonic stages.抄録全体を表示PDF形式でダウンロード (4176K) -
Masao Iwamori, Motoko Sakayori, Shirou Nozawa, Tomotaka Yamamoto, Masa ...1989 年 105 巻 5 号 p. 718-722
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーA monoclonal antibody, MSN-1, generated by immunizing a mouse with a human uterine endometrial adenocarcinoma cell line, SNG-II, was strongly and specifically reactive with neutral glycosphingolipids from cancer tissues of patients with uterine endometrial adenocarcinomas. The glycosphingolipid antigen was purified from pooled human meconia, which contained the antigen at the concentration of 1.95 μmol/g dry weight. Its structure was determined by NMR, negative ion FABMS, permethylation analysis, and TLC-immunostaining with monoclonal anti-Lc4, Cer antibody, and was concluded to be the III4IV2Fuc2Lc4, Cer, Leb antigen of the human Lewis blood system. On ELISA, the monoclonal antibody was found to be strongly reactive with Leb, slightly with Lea and not at all with A, B, H, or IV2FucGg4Cer. The amount of Leb in cancerous regions in the patients with the Lea-b+ blood group was significantly increased compared to that in normal regions in the same patients, and it was a newly appearing antigen in the cancerous tissue of a patient with the Leaa+b- blood group.抄録全体を表示PDF形式でダウンロード (3009K) -
Masao Iwamori, Tatsuya Kawaguchi, Yoshitaka Nagai1989 年 105 巻 5 号 p. 723-727
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーNeuraminidase treatment of lymphocytes is known to cause changes of cellular responses in several biological phenomena, but the molecules modified on the cell surface by neuraminidase are not known in detail. Rabbit thymocytes, which contain tissue-characteristic gangliosides, were treated with Vibrio cholerae neuraminidase, and the susceptibility of the cell surface sialic acid residues was examined. The amount of sialic acid released from the thymocytes at the highest level was 42.4nmol per 1×109 cells, among which 26.5% was from gangliosides. Ninety-three percent of the VI3NeuGc-nLc6Cer, 84% of the IV3NeuGc-nLc4Cer, and 50% of the II3NA2-LacCer in the thymocytes was hydrolyzed to nLc6Cer, nLc4Cer, and LacCer, respectively, but II3NA-LacCer was completely cryptic. Also, among the molecular species of II3NA2-LacCer, C20:0- to C24:0-containing, but not C16:0- to C18:0-containing molecules, were susceptible to neuraminidase. After neuraminidase treatment, nLc4Cer and nLc6Cer became the major glycosphingolipids, and a 15-fold increase of radioactivity incorporated into the glycosphingolipids was observed by the galactose oxidase-sodium borotritide procedure, suggesting that the β-galactose of the glycosphingolipids produced by neuraminidase treatment is accessibly to the several ligands which are functionally associated with lymphocytes.抄録全体を表示PDF形式でダウンロード (3103K) -
Katsuko Yamashita, Kazuhide Totani, Yo Iwaki, Ikuko Takamisawa, Noriko ...1989 年 105 巻 5 号 p. 728-735
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーThe sugar chains of γ-glutamyltranspeptidases, purified from human hepatoma and from normal human liver, were released quantitatively as oligosaccharides by hydrazinolysis. Comparative study of their structures revealed that the following structural alterations are induced by hepatocyte carcinogenesis: 1) high mannose type sugar chains are detected in the hepatoma enzyme but not in the normal liver enzyme; 2) abnormal biantennary sugar chains containing C-2, 4 outer chain branches newly appeared; 3) the total amounts of tri- and tetraantennary sugar chains containing C-2, 6 outer chain branches increased up to three times.抄録全体を表示PDF形式でダウンロード (918K) -
Kazuko Kita, Toshio Otsuki, Toshiharu Ishizuka, Masamiti Tatibana1989 年 105 巻 5 号 p. 736-741
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーMammalian phosphoribosyl pyrophosphate (PRPP) synthetase has been extensively investigated. However, considerable ambiguity remains concerning its physical and regulatory properties. We purified PRPP synthetase from rat liver and studied some of the physical properties, in parallel with cloning experiments (Taira, M. et al. [1987] J. Biol. Chem. 262, 14867-14870). 1) The enzyme was purified to a specific activity of 7, 280 milliunits/mg, the highest value in the literature for a mammalian PRPP synthetase. The apparent molecular mass was over 1, 000kDa. 2) The final preparation contained 34-kDa, 38-kDa, and 40-kDa protein species, as analyzed by SDS gel electrophoresis. 3) Further attempts at separation using conventional procedures only led to a co-purification of the components. Thus, the purified enzyme appears to exist as complex aggregates composed of heterogeneous components. 4) Gel filtration of the enzyme in the presence of 1M MgCl2 isolated part of the 34-kDa component, free of other species. The preparation was catalytically active, indicating that this component is the catalytic subunit. 5) The amino acid composition of the 34-kDa subunit and the amino acid sequences of its N-terminal region and of two tryptic peptides were determined. The results are in accord with the results of cDNA analyses.抄録全体を表示PDF形式でダウンロード (2919K) -
Masaya Takamoto, Tamao Endo, Mamoru Isemura, Naohisa Kochibe, Akira Ko ...1989 年 105 巻 5 号 p. 742-750
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーThe asparagine-linked sugar chains of fibronectin purified from human placenta were quantitatively released as oligosaccharides by hydrazinolysis. After N-acetylation, they were converted to radioactive oligosaccharides by NaB3H4 reduction. The radioactive oligosaccharides were fractionated by their charge on an anion-exchange column chromatography. All of the acidic oligosaccharides could be converted to neutral oligosaccharides by sialidase digestion. These oligosaccharides were then fractionated by serial affinity chromatography using immobilized lectin columns. Study of each oligosaccharide by sequential exoglycosidase digestion and methylation analysis revealed the following information as to the structures of the sugar chains of human placental fibronectin: 1) nine sugar chains are included in one molecule; 2) all sialic acid residues are exclusively linked at the C-3 position of the galactose residues; 3) bi-, tri-, and tetraantennary complex-type oligosaccharides with the Manα1→6(Manα1→3)Manβ1→4GlcNAcβ1→4(±Fucα1→6)-GlcNAc as their cores were found; 4) the bisecting N-acetylglucosamine residue and the Galβ1→4G1cNAcβ1→ repeating groups are included in some of the sugar chains.抄録全体を表示PDF形式でダウンロード (1125K) -
Hiroshi Funabashi, Akihiko Kawaguchi, Hiroshi Tomoda, Satoshi Omura, S ...1989 年 105 巻 5 号 p. 751-755
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーAn antibiotic cerulenin, (2R, 3S)-2, 3-epoxy-4-oxo-7, 10-trans, trans-dodecadienamide, irreversibly inhibits fatty acid synthetase from Saccharomyces cerevisiae. Three moles of cerulenin were bound to 1mol of the enzyme with concomitant loss of its activity. Pretreatment of the enzyme with iodoacetamide reduced the amount of cerulenin bound to the enzyme. Since iodoacetamide is known to specifically bind to the cysteine residue on the condensing reaction domain, cerulenin is considered to bind to the same domain. Tryptic digestion of the [3H]cerulenin-treated enzyme gave a radioactive peptide; its amino acid composition was Asx 1, Thr 1, Ser 1, Glx 2, Pro 1, Gly 1, Ala 1, Val 1, Ile 1, and Leu 2. This composition included all the amino acids of the condensing reaction site (Thr-Pro-Val-Gly-Ala-Cys) previously reported by Kresze et al. (Eur. J. Biochem., 79, 181 [1977]) except for Cys. When the enzyme was treated with [3H]cerulenin and digested successively with trypsin and carboxypeptidase P, a [3H]cerulenin-cysteine adduct was isolated as the sole product. This was identified with the adduct chemically synthesized from non-labeled cerulenin and cysteine, and its structure was elucidated by 1H-, 13C-NMR, and fast atom bombardment mass spectrometry. These results indicate that cerulenin, forming a hydroxylactam ring, reacts at its epoxide carbon (C-2 position) with the SH-group of the cysteine residue in the condensing reaction domain of yeast fatty acid synthetase.抄録全体を表示PDF形式でダウンロード (539K) -
Toshiyuki Sakai, Akira Yoshioka, Kazukuni Yamamoto, Koh Niinomi, Yoshi ...1989 年 105 巻 5 号 p. 756-759
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーHemophilia B Kashihara is a severe hemorrhagic disorder in which the factor IX antigen is present in normal amounts but factor IX biological activity is markedly reduced. In addition, purified factor IX Kashihara is not activated by purified factor XIa in the presence of calcium ions. Amino acid sequence analysis of one of the tryptic peptides isolated from factor IX Kashihara indicated that Val-182 (equivalent to Val-17 in the chymotrypsin numbering system) had been replaced by Phe. No substitution was found in the members of the catalytic triad His-221, Asp-269, and Ser-365 of factor IX Kashihara. The Val-to-Phe replacement found in factor IX Kashihara appears to sterically hinder the cleavage of Arg 180-Val 181 by factor XIa required for the activation of this zymogen.抄録全体を表示PDF形式でダウンロード (518K) -
Seiji Ichida, Takashi Fujisue, Atuhiro Masada, Yasuo Oda, Narihisa Mat ...1989 年 105 巻 5 号 p. 760-766
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーThe characteristics of the specific bindings of [3H] nitrendipine (Nit) and [3H] (+)PN200-110 (PN) to crude membranes from rat skeletal, cardiac, and uterine muscle and whole brain were investigated, with special interest in the effect of UV irradiation on these bindings. The specific bindings of [3H]Nit and [3H](+)PN to these crude membranes were saturable and reversible. The specific bindings of [3H]Nit to all these membranes except crude skeletal membranes was maximum in the presence of 0.15 M NaCI plus 1 mM CaC12 and minimal in the absence of these ions, but the specific bindings of [3H](+)PN to these crude membranes was not affected significantly by these ions. A calcium agonist and antagonists inhibited the specific bindings of [3H]Nit and [3H](+)PN to these crude membranes, the order of their inhibitory effects on specific [3H]Nit bindings being roughly Nit _??_(+)PN_??_(-)PN_??_Bay K 8644 (Bay)> verapamil (Ver)> diltiazem (Dil). In crude skeletal membranes only, PN caused significant stereospecific inhibition. The order of inhibitions of specific [3H] (+)PN bindings to these crude membranes was generally (+)PN > Nit _??_ (-)PN > Bay_??_ Ver_??_Dil. In all these crude membranes, UV irradiation completely prevented decrease in the amount of specific binding of [3H]Nit on addition of a large excess of unlabeled Nit and reduced the decrease of specific [3H] (+)PN binding on addition of excess unlabeled (+)PN. These findings suggested that [3H]Nit and [3H](+)PN bind to voltage-sensitive calcium channels in crude membranes from rat skeletal, cardiac, and uterine muscle and whole brain, and that UV irradiation changes the specific bindings of [3H] Nit and [3H] (+)PN from reversible to irreversible bindings.抄録全体を表示PDF形式でダウンロード (723K) -
Seiji Ichida, Atsuhiro Masada, Takashi Fujisue, Takuya Yoshioka, Narih ...1989 年 105 巻 5 号 p. 767-774
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーThe characteristics of photoaffinity labeling with the calcium agonist [3H]Bay K 8644 (Bay) and the calcium antagonists [3H] nitrendipine (Nit) and (+)PN200-110 (PN) of crude membranes from rat skeletal, cardiac, Heal, and uterine muscles and whole brain were investigated. In all these crude membranes, [3H] (+)PN (20 nM) was mainly photoincorpo-rated into one protein band with a molecular weight of 30, 000 - 41, 000 Da. It was also incorporated into some other bands of all these crude membranes. The photoincorporation of [3H] (+)PN into these crude membranes was inhibited by the presence of 20 μM unlabeled (+)PN. The photoincorporation of [3H] (+)PN into these crude membranes depended on its dose and on the time of UV irradiation. No incorporation of [3H] (+)PN was observed in the absence of UV irradiation. The incorporation was not affected by the presence of 1 mM CaC12 and/or 0.15 M NaC1, but was significantly decreased by 20 μM (+)PN and slightly de-creased by 20 μM (-)PN, 20 μM Bay, 1mM diltiazem, or 1mM verapamil. Namely, enantiomers of PN caused various extents of stereoselective inhibition of photoaffinity labeling by [3H] (+)PN of specific protein bands in these crude membranes. [3li]Nit was photoincorporated into these crude membranes in the same way as [3H] (+)PN, but [3H] Bay was not photoincorporated. However, 20 μM unlabeled Nit did not consistently inhibit photoaffinity labeling with [3H] Nit. These findings suggested that measurement of photoaffinity of crude membranes from rat skeletal, cardiac, and uterine muscles and whole brain with [3H] (+)PN by UV irradiation is a useful method for investigating the character-istics of the voltage-dependent calcium channels that are affected by 1, 4-dihydropyridine derivatives.抄録全体を表示PDF形式でダウンロード (4910K) -
Yasutada Imamura, Masao Kawakita1989 年 105 巻 5 号 p. 775-781
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーSarcoplasmic reticulum membranes were treated with trypsin, and samples enriched with A1a, A1b, and C fragments (Saito, K. et al. (1984) J. Biochem. 95, 1297-1304), respectively, were prepared. A1b, and C fragments were purified to apparent homogeneity, and an approximately equimolar mixture of A1(Met1-Arg198), A1a, and A1b fragments free from other contaminants was also obtained through gel permeation and hydroxylapatite chromatography in the presence of sodium dodecyl sulfate. N- and C-terminal amino acid sequence analyses of these peptid es were carried out in order to identify the tryptic cleavage sites responsible for the formation of these fragments. Both A1a and A1b fragments had the same C-terminal sequence as A1 fragment. Single cleavage of A1 at T3a (Lys218-A1a219) yielded A1a, while a cleavage between either Lys234-11e235 or Arg236-Asp237 (collectively designated as T3b) resulted in Alb fragment. Thus, A1a and A1b fragments differed from A1 fragment only by their loss of short stretches corresponding to the N-terminal region of the latter. On the other hand, C fragment represented the C-terminal half of B fragment (Ala506- Gly994). It had the same C-terminal sequence as B fragment and was produced by cleavage at T4 (Lys728-Thr729). Cleavages at T3a and T3b profoundly affected the catalytic properties of SR-ATPase (Imamura, Y. and Kawakita, M. (1986) J. Biochem. 100, 133-141), and it was suggested that the segment of the ATPase molecule including the region between Ala199 and Arg236 is important in mediating the coupling between ATP splitting and Ca2+-transport.抄録全体を表示PDF形式でダウンロード (796K) -
Michinao Mizugaki, Michiko Nakazawa, Hirotaka Yamamoto, Hiroshi Yamana ...1989 年 105 巻 5 号 p. 782-784
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーN-Ethylmaleimide (NEM) reducing enzyme was purified to homogeneity from cell-free extracts of Candida lipolytica by chromatographic techniques. The molecular weight of the native enzyme was estimated to be about 43, 000 by gel filtration using Superose 12TM and to be 47, 000 by SDS-PAGE. This enzyme can use both NADPH and NADH as an electron donor, and catalyzes the reduction of the carbon-carbon double bond of five membered ring compounds which have two conjugated carbonyl groups on both sides of a double bond.抄録全体を表示PDF形式でダウンロード (325K) -
Hiroko Itami, Yuki Sakai, Tadashi Shimamoto, Hiroko Hama, Masaaki Tsud ...1989 年 105 巻 5 号 p. 785-789
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーMembrane-bound 5'-nucleotidase from Vibrio parahaemolyticus was solubilized and purified using a nonionic detergent, heptyl-β-D-thioglucoside, and was characterized. This enzyme required Mg2+ for activity, maximum activity being observed at 5 and 20 mM Mg2+ with AMP and ATP, respectively, as substrates. Of the divalent cations tested, Mn2+ and Co2+ were able to replace Mg2+ partially, whereas Ca2+ was ineffective. Zinc strongly inhibited the enzyme activity and Ni2+ caused partial inhibition. This enzyme required Cl-for activity, the optimal concentration being 20 mM or more. The order of effectiveness of anions was Cl- > Br- >I- ≈ NO3-. Sulfate and acetate were ineffective. The optimal pH was 8.0. The activity of the purified enzyme was stimulated by the addition of lipid to the assay mixture. This enzyme hydrolyzed all 5'-nucleotides tested, but did not hydrolyze 3'-nucleotides or ribose 5-phosphate. On sodium dodecyl sulfate-polyacrylamide gel electro-phoresis, the enzyme appeared to be a single polypeptide, with a molecular weight of 72 kDa.抄録全体を表示PDF形式でダウンロード (575K) -
Koshi Saito, Kunihiro Yamamotc, Iwao Nagayama, Junko Uemura, Takeshi K ...1989 年 105 巻 5 号 p. 790-793
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーNa+-dependent I- transport and I- counterflow were studied using phospholipid vesicles (P-vesicles) made of porcine thyroid plasma membranes and soybean phospholipid by sonication. 1) I- uptake by P-vesicles incubated in the presence of external Na+ was higher than that by P-vesicles incubated in choline+ instead of Na+. The vesicles exhibited Na+-dependent I- uptake. When P-vesicles were internally loaded with I- prior to incuba-tion in Na+, a further increase in Na+-dependent I- uptake was observed, although the concentration of internal I- was very much higher than that outside. In the absence of external Na+, I- uptake by P-vesicles preloaded with I- was comparable to baseline uptake. 2) Na+-dependent I- uptake by P-vesicles not loaded with I- and enhanced Na+-dependent I- uptake by P-vesicles preloaded with I- were both inhibited by either of SCN- and ClO4-added outside the vesicles. 3) When P-vesicles were loaded with SCN- instead of I- and incubated in Na+, I- uptake by these vesicles was also higher than baseline Na+-dependent I- uptake. However, a ClO4- load did not result in an increase in I- uptake. These results indicate that Na+-dependent I- transport including Na+ -dependent I- counterflow is specifically mediated by the thyroid I- carrier. SCN--I- counterflow in addition to I--I-counterflow occurs dependently on Na+, but ClO4--I- counterflow does not.抄録全体を表示PDF形式でダウンロード (454K) -
Hiroshi Yoshida, Shigeo Oikawa, Megumi Ikeda, Elwyn T. Reese1989 年 105 巻 5 号 p. 794-798
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーAn acid phosphatase has been purified from the culture broth of Penicillium funiculosum by procedures including SP-Sephadex column chromatography and Sephacryl S-200 gel filtration. The phosphatase appears to be a 76 kDa heterodimer composed of 51 and 26 kDa subunits on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme hydrolyzes both phosphodiesters and phosphomonoesters, but only those with aryl leaving groups. At the early phase of degradation of bis-p-nitrophenyl phosphate by the enzyme, inorganic phosphate and the intermediary product, p-nitrophenyl phosphate, are liberated at approximately the same rate. This indicates that the intermediary phosphomonoester produced on the enzyme is further hydrolyzed in situ or dissociates into the medium at approximately the same probability.抄録全体を表示PDF形式でダウンロード (1427K) -
Hideo Yamamoto, Minoru Terabayashi, Takanobu Egawa, Eijiro Hayashi, Hi ...1989 年 105 巻 5 号 p. 799-802
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーHuman plasma gelsolin was specifically eluted with 1 mM adenosine 5' -triphosphate from an Affi-Gel Blue column. Since the ionic strength of sodium chloride required to elute the protein from the dye column was much higher than that of 1 mM adenosine 5'-triphosphate, the binding of plasma gelsolin with the dye-ligand appeared to be biospecific. Taking advantage of this affinity interaction, we have developed a revised purification method of human plasma gelsolin. The purification included ammonium sulfate precipitation, diethylaminoethyl-Sepharose chromatography, Affi-Gel Blue chromatography, and Phenyl-Sepharose chromatography. The method allowed a reproducible purification of the protein to apparent homogeneity, producing a 331-fold purification with a yield of 6%.抄録全体を表示PDF形式でダウンロード (2594K) -
Sueo Matsumura, Keizo Ohmori, Tanemichi Chiba, Akira Kumon1989 年 105 巻 5 号 p. 803-812
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーAn antibody obtained by immunizing a rabbit with purified bovine brain myosin was found to react with the tail portion of the myosin heavy chain. An Fab fragment obtained by limited papain digestion of the antibody was allowed to bind to brain myosin, and the complex of the Fab fragment and brain myosin (Fab-myosin.) was isolated. On examination of the rotary-shadowed Fab-myosin by electron microscopy, most of the Fab fragment was located on the middle to C-terminal regions of the tails of the myosin molecules. The solubility of Fab-myosin in low salt solutions was higher than that of control brain myosin. Fab-myosin was found to form small irregular aggregates in low salt solutions instead of regular bipolar filaments, and the relative population of the monomeric form of myosin molecules observed for the Fab-myosin was much larger than that observed for the control myosin. The actin-activated Mg2+-ATPase activity of Fab-myosin was stimulated two- to threefold by phosphorylation of the light chains with myosin light chain kinase, as observed for the control brain myosin. Furthermore, the levels of the ATPase activity of the phosphorylated and dephosphorylated Fab-myosins were similar to those of the phospho-rylated and dephosphorylated control myosins, respectively. The superprecipitation activity of Fab-myosin was also highly dependent on phosphorylation of the light chains. Although control brain myosin formed a large superprecipitate network which contracted to a dense particle, Fab-myosin generated only numerous tiny superprecipitates under the same conditions. From these results it was deduced that a regular filamentous state of brain myosin was not prerequisite for its actin-activated Mg2+-ATPase and superprecipitation activities but was indispensable for the formation of a large and well contractible super-precipitate.抄録全体を表示PDF形式でダウンロード (6784K) -
Akira Taketo1989 年 105 巻 5 号 p. 813-817
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーEfficient and reproducible DNA-transfection was attained in E. coli, by electroporation. The yield of the transfectants was affected by pretreatment of the recipient cells as well as by the composition of the electroporation medium. Using a single pulse procedure, relationships among the electrical parameters, the transfection efficiency, and the cellular viability were investigated in 10 mM Tris-HC1 buffer (pH 7. 5) containing 5% sucrose. Certain sodium salts (e. g. citrate, phosphate, and sulfate) were promotive, whereas Mg2+, DEAE-dextran, and polyvinylpyrrolidone were inhibitory to the transfection. Heter-ologous nucleic acids (native DNA, denatured DNA, and tRNA.) exerted only a marginal effect on transfection with a viral replicative-form DNA. The efficiency of DNA transfer was affected by culture conditions, and bacteria grown at a higher temperature were more competent. The electroporation system was more efficient than an improved CaC12 method, not only in transfection with viral single- and double-stranded DNAs, but also in transfor-mation with plasmid DNAs.抄録全体を表示PDF形式でダウンロード (586K) -
Toshio Ando1989 年 105 巻 5 号 p. 818-822
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーF-Actin was partially cross-linked to myosin subfragment-1 (S-1) at various molar ratios (r=S-1/actin) with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. The cross-linked acto-S-1 ATPase showed so called “super-activation, ” VX. S-1 was added further to the cross-linked acto-S-1 and the ATPase activity, VY, was measured. Since the added S-1 can interact only with the bare actin protomers within the cross-linked actin filament, the difference, ΔV=VY-VX-VS, (where VS is the ATPase activity of the additional S-1 alone), can indicate the state of the bare actin protomers while the cross-linked acto-S-1 is hydrolyzing ATP. With increasing r, ΔV decreased much more rapidly than ΔV0(1-r) (where ΔV0. is ΔV at r=0.) and reached a minimum around r=0.15. As r increased further, , ΔV approached the level of ΔV0(1-r). When Sil1/SH2-blocked S-1 was cross-linked to F-actin, ΔV decreased according to ΔV0(1-r). Therefore, the large reduction of ΔV, observed when intact S-1 was cross-linked, was coupled to the high ATPase activity of the cross-linked acto-S-1. Combining these data with other kinetic data, we could deduce that structural distortion in a cross-linked actin induced by the ATPase reaction of the S-1 partner propagated over several bare actin protomers along the filament and reduced their affinity for the S-1-ADP-P1 complex. A model is presented to interpret these data, hypothe-sizing an asymmetry in the propagation of actin conformational change.抄録全体を表示PDF形式でダウンロード (578K) -
Tomoyoshi Kobayashi, Osamu Kagami, Takashi Takagi, Kazuhiko Konishi1989 年 105 巻 5 号 p. 823-828
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーThe amino acid sequence of troponin C obtained from horseshoe crab, Tachypleus tridentatus, striated muscle was determined by sequence analysis and alignments of chemically and enzymatically cleaved peptides., Troponin C is composed of 153 amino acid residues with a blocked N-terminus and contains no tryptophan or cysteine residue., The site I, one of the four Ca2+-binding sites, is considered to have lost its ability to bind Ca2+ owing to the replacements of certain amino acid residues.抄録全体を表示PDF形式でダウンロード (540K) -
Minoru Suzuki, Michiko Sekine, Tamio Yamakawa, Akemi Suzuki1989 年 105 巻 5 号 p. 829-833
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーA method of high-performance liquid chromatography-fast atom bombardment mass spectrometry (HPLC/FAB/MS) for the structural characterization of glycosphingolipids was developed, which involves a frit interface between the HPLC and the MS. The molecular species of glucosylceramide (GlcCer) purified from the spleen of a patient with Gaucher's disease and galactosylglobotetraosylceramide (IV3βGal-Gb4Cer) from mouse kidney were analyzed using this system on a reversed-phase column, with methanol containing 1% glycerol as the elution solvent. The injection of 1μg of GlcCer gave the mass spectra of seven major molecular species, the pseudo-molecular ion for each of the seven molecular species being observed at m/z 698, 726, 754, 782, 808, 796, and 810, respectively. The injection of 200 pg of synthetic N-stearoyl glucosylsphingosine (d18:1) gave a clear peak with the single ion monitoring method detecting the pseudo-molecular ion at m/z 726. The injection of 5μg of IV3β Gal-Gb4Cer gave the mass spectra of six major molecular species, the pseudo-molecular ions being observed at m/z 1, 489, 1, 471, 1, 515, 1, 497, 1, 517, and 1, 499. This report deals with a new HPLC/FAB/MS system, which was successfully applied to the structural characterization of the molecular species of neutral glycosphingolipids, and the system is a quite promising for development into a quantitative method for glycosphingolipids with high sensitivity and specificity.抄録全体を表示PDF形式でダウンロード (486K) -
Hiroyasu Inoue, Kiyoshi Fukui, Yoshihiro Miyake1989 年 105 巻 5 号 p. 834-840
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーGenomic clones carrying the entire rat true tissue kallikrein gene have been isolated and characterized. Southern blot analysis revealed the presence of a unique gene for true tissue kallikrein in the rat haploid genome. The true tissue kallikrein gene was composed of 5 exons and 4 introns, its length being about 4.5 kilobase pairs. We also isolated the rat renal kallikrein cDNA which was expressed in the kidney. The exon sequences of the gene were identical with the corresponding sequences of the kallikrein cDNA. The rat true tissue kallikrein gene was distinct from two rat kallikrein-related genes expressed in the kidney (Chen, Y. et al. (1988) Biochemistry 27, 7189-7196). Primer extension analysis indicated that two identical transcription-initiation sites were used in both the kidney and subman-dibular gland, although expression of the true tissue kallikrein in the submandibular gland was much higher than that in the kidney. The 3'-noncoding region of this gene contained the polyadenylation signal, AATAAA, observed in the rat renal kallikrein cDNA. The 5'-flanking region was sequenced up to about 800 base pairs, upstream of the cap sites. CATCT, GGGCGG, and TTTAAA boxes were found as common signals required for eukaryote gene expression within about 100 base pairs-upstream of the cap sites.抄録全体を表示PDF形式でダウンロード (2961K) -
Yuki Sakai, Yumi Tamao, Tadashi Shimamoto, Hiroko Hama, Masaaki Tsuda, ...1989 年 105 巻 5 号 p. 841-846
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーThe gene encoding the membrane-bound 5'-nucleotidase of Vibrio parahaemolyticus was cloned and expressed in Escherichia coli. Cells of E. coli harboring a plasmid, pNUT5, which carries the 5'-nucleotidase gene were able to grow on ATP as the sole source of carbon, although the original cells were not. The 5' -nucleotidase activity was detected in whole cells of E. coli harboring pNUT5 and in membrane vesicles prepared from these cells. Most properties of the 5'-nucleotidase produced in E. coli, that is, its requirements for Cl- and Mg2+, substrate specificity, and inhibition by Zn2+, were similar to those observed in V. parahaemolyticus, but some alterations in properties were observed: The 5'- nucleotidase was partially inducible in V. parahaemolyticus, but its expression in E. coli was completely constitutive. The specific activity of the 5'-nucleotidase in membrane vesicles of E. coli harboring the plasmid was 30 times that observed in whole cells, whereas the specific activities in membrane vesicles and in whole cells of V. parahaemolyticus were almost the same. A new, dense band of protein with an apparent molecular mass of 63 kDa was detected when membrane proteins of E. coli harboring the plasmid were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.抄録全体を表示PDF形式でダウンロード (1785K) -
Toshiyuki Miyata, Hiroyuki Takeya, Yasushi Ozeki, Mitsue Arakawa, Fumi ...1989 年 105 巻 5 号 p. 847-853
発行日: 1989年
公開日: 2008/11/18
ジャーナル フリーThe complete amino acid sequence and disulfide bridge location of HR2a, one of the hemorrhagic proteins isolated from the snake venom of Trimeresurus flavoviridis, have been determined by analysis of peptides derived from digests with cyanogen bromide, lysyl endopeptidase, trypsin, and Staphylococcus aureus V8 protease. Peptides were purified by gel filtration followed by reversed-phase HPLC. HR2a has the amino-terminal sequence of < Glu-Gln-Arg- and consists of a total of 202 residues with a calculated molecular weight of 23, 015. Sequence analysis indicates the presence of another isoform which lacks the amino-terminal residue, making 201 amino acid residues with a molecular weight of 22, 887. Three disulfide bridges of HR2a link Cys-118 to Cys-197, Cys-159 to Cys-181, and Cys-161 to Cys-164. HR2a contains a segment which is similar to the zinc-chelating sequences found in thermolysin and several mammalian metalloproteinases, suggesting that HR2a is a metalloproteinase with limited substrate specificity. However, there is no other significant sequence homology with thermolysin except for the zinc-ligand region.抄録全体を表示PDF形式でダウンロード (830K)
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