巻号一覧
90 巻, 5 号
選択された号の論文の43件中1～43を表示しています
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Kazuo ASAOKA, Kenji TAKAHASHI1981 年 90 巻 5 号 p. 1237-1242
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーA simple and sensitive enzymatic assay method for the determination of reduced glutathione (GSH) has been developed using glutathione S-aryltransferase with o-dinitrobenzene as a substrate. o-Dinitrobenzene is a good substrate for the enzyme and has low spontaneous reactivity with GSH at neutral pH. GSH can be determined by colorimetrically measuring nitrite released upon the enzymatic conjugation of GSH and o-dinitrobenzene by using a diazo-coupling method with N-(1-naphthyl)ethylenediamine dihydrochloride. This method is capable of quantitating 1 to 40 nmol of GSH, and can be applied to physiological samples containing deproteinizing reagents.抄録全体を表示PDF形式でダウンロード (406K) -
Kazuhiro SOGAWA, Yoshikazu ICHIHARA, Kenji TAKAHASHI1981 年 90 巻 5 号 p. 1243-1248
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーSome characteristics of the membrane-bound neutral proteinase activity in the microsomal fractions of rat kidney and small intestine were compared, using heatdenatured casein as a substrate. The proteinases of both kidney and small intestine showed maximal activity between pH 8.0 and 8.5, and were strongly inhibited by EDTA, o-phenanthroline, p-chloromercuriphenyl sulfonate, dithiothreitol, and chymostatin. Phosphoramidon and other reagents tested, including deoxycholate and bestatin, which strongly inhibited the contaminating aminopeptidase activity, were without marked effect on the neutral proteinase activity. Among ureadenatured proteins tested as substrates, casein, histone, and hemoglobin were hydrolyzed rapidly by both proteinase preparations. Fibrinogen was a good substrate for the kidney enzyme whereas it was not hydrolyzed well by the small intestine proteinase. On the other hand, serum albumin was hydrolyzed well by the small intestine proteinase, but not by the kidney proteinase. These results indicate that the neutral proteinase activity of the microsomal membrane fractions is largely due to metalloproteinases, which are quite similar, but not identical, in the kidney and small intestine.抄録全体を表示PDF形式でダウンロード (413K) -
Noriko FUJII, Yutaka NAGAI1981 年 90 巻 5 号 p. 1249-1258
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーA proteodermatan sulfate was extracted from calf skin with 3M MgCl2 at 4°C in the presence of protease inhibitors and purified by DEAE-cellulose chromatography followed by CsCI density gradient centrifugation. The molecular weight of the proteoglycan was estimated to be approximately 115, 000, based on the results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
The proteoglycan contains neutral sugars such as galactose, xylose, mannose, fucose, and glucose, but its amino acid composition is similar to that of proteodermatan sulfates from bovine heart valve, tendon, sclera, and pig and rat skin. The core protein is a glycoprotein with a molecular weight of 56, 000 and seems to be monodisperse. However, the dermatan sulfate component, which is the only glycosaminoglycan constituent composed of 95% 4-sulfated disaccharide unit and 5% 6-sulfated disaccharide unit, seems to be polydisperse with a number average molecular weight of 17, 000, as judged from both gel chromatography and chemical analysis. Analysis by β-elimination and reduction of the dermatan sulfate-peptide(s) indicated that the glycan components are linked to the core protein through an O-glycosidic linkage between xylose and serine residues.
From these results, we propose a model for the structure of calf skin proteodermatan sulfate that is composed of 3-4 chains of dermatan sulfate components which are covalently linked to a core (glyco-) protein via O-glycosidic linkages.抄録全体を表示PDF形式でダウンロード (1565K) -
Kimitsuna WATANABE, Keiko ASAI, Tairo OSHIMA, Yoshiyuki KUCHINO1981 年 90 巻 5 号 p. 1259-1266
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe nucleotide sequence of initiator tRNA (tRNAiMet) from Euphausia sperba, which was harvested in the Antarctic Sea, was determined to be pA-G-C-A-G-A-G-U-m1G-m2G-C-G-C-A-G-U-G-G-A-A-G-C-G-U-m2G-C-U-G-G-G-C-C-C-A-U-t6A-A-C-C-C-A-G-A-G-m7G-U-C-G-G-U-A-G-A-ΨC-G-m1A-A-A-C-U-A-C- U-C-U-C-U-G-C-U-A-C-C-AOH by using post-labeling methods recently developed. The nucleotide sequence was very similar to that of mammalian tRNAiMet except for changes in six bases and three modifications: C16, U55, D47 and m5C48 are replaced by U16, Ψ55 and unmodified U47 and C48, respectively. A50-U64 and G52-C62 base pairs of mammalian tRNAimet are reversed in Euphausia tRNAiMet. In addition, the G49-C65 pair of the former is replaced by a less stable G49-U65 pair in Euphausia tRNAiMet. The sequence homology was compared between Euphausia tRNAiMet and over ten different species of eucaryotic tRNAiMet so far sequenced. The melting temperature of Euphausia tRNAiMet was 72.5°C, which is 4.2°C and 8.3°C lower than those of rat liver and yeast tRNAiMet, respectively. The origin of the thermal instability of Euphausia tRNAiMet is discussed in comparison of its secondary structure compared with those of other eucaryotic tRNAiMet.抄録全体を表示PDF形式でダウンロード (1062K) -
Tarou OGURUSU, Akio MAEDA, Norihiro SASAKI, Tôru YOSHIZAWA1981 年 90 巻 5 号 p. 1267-1273
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリー1. Membrane fraction containing halorhodopsin was prepared from the lysate of a mutant strain of Halobacterium halobium, Y1, which is defective in bacteriorhodopsin synthesis.
2. Irradiation of the membrane with red light at 0°C decreased the absorbance intensities over the whole range of 500-600 nm and a new absorption peak appeared at about 400 nm. This process was reversed either by irradiation with blue light or simply by incubation in the dark. The wavelength at which the red light-induced absorbance decrease became maximum was 566 nm in the absence of NaCl and moved to 576 nm upon addition of NaCl. The midpoint for the conversion was at about 0.1M NaCl.
3. Even though the membrane containing halorhodopsin was prepared in the absence of NaCl, this pigment was photochemically active in the sense that it could form a bathochromic photoproduct, a batho-intermediate, at 196°C.
4. Retinal isomer composition during irradiation with red light at 0°C was determined. Unirradiated halorhodopsin had predominantly all-trans retinal. With the increase of the blue-shifted product on irradiation, 13-cis content increased.抄録全体を表示PDF形式でダウンロード (567K) -
Sumihiro HASE, Tokuji IKENAKA, Yoshio MATSUSHIMA1981 年 90 巻 5 号 p. 1275-1279
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe electrophoretic mobilities of pyridylamino derivatives of oligosaccharides in paper electrophoresis at pH 5 ((1979) J. Biochem. 85, 989-994) were independent of linkage positions, anomeric configurations, and acetamido groups up to a molecular weight of 1, 700 and dependent on only their molecular weights. To increase the accuracy and sensitivity of the method, the pyridylamino derivative of a sample (10-50 pmol) was mixed with small amounts of pyridylamino derivatives of internal standard oligosaccharides of different molecular weights. The mixture was electrophoresed on a paper. Sections of paper corresponding to the standard oligosaccharides were extracted with water and then each extract was analyzed by high performance liquid chromatography with a fluorescence spectrophotometer. The molecular weight of the pyridylamino derivative of the sample was calculated by comparing its mobility with those of the pyridylamino derivatives of the internal standard oligosaccharides which were detected together with the sample on the chromatogram.抄録全体を表示PDF形式でダウンロード (1066K) -
Katsuko YAMASHITA, Yoko TACHIBANA, Toshiyuki TAKEUCHI1981 年 90 巻 5 号 p. 1281-1289
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーAbout thirty percent of two α-amylases produced from a serous papillary cystadenocarcinoma of the ovarium (case 1) and a bronchioloalveolar adenocarcinoma of the lung (case 2) was glycoproteins containing 1 mol of asparagine-linked sugar chain, respectively. The structures of the sugar moieties were found by sequential enzymatic degradation and methylation analysis to be as follows: [(Galβ1→4)0or 1GlcNAcβ1→2Manα1→6(3)] [GlcNAcβ1→2Manα1→3(6)] Manβ1→4GIcNAcβ1→4(Fucα1→6)GlcNAc and [(Galβ→4)0or1GlcNAcβ2Manα→1] [NeuAcα2→6Galβ1→4GlcNAcβ1→2Manα1-3] Manβ1→4G lcNAcβ1→4(Fucα1→6)GlcNAc. Structures of asparagine-linked sugar chains were the same in the tumors of cases 1 and 2 and were incomplete in comparison with those of the parotid amylase.抄録全体を表示PDF形式でダウンロード (701K) -
Yoko TACHIBANA, Katsuko YAMASHITA, Masao KAWAGUCHI, Shin-ichiro ARASHI ...1981 年 90 巻 5 号 p. 1291-1296
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe substrate specificity of human endo-β-N-acetylglucosaminidase was studied by using the homogenate of cultured skin fibroblasts of fucosidosis patients as an enzyme source. The results indicate that biantennary complex type asparaginelinked sugar chains as well as high mannose type sugar chains are cleaved by the enzyme action. None of the sugar chains with a fucosyl residue on the proximal N-acetylglucosamine of their N, N'-diacetylchitobiose moieties was cleaved. These results proved enzymatically the mechanism of production of oligosaccharides detected in the urine of various exoglycosidase deficiencies.抄録全体を表示PDF形式でダウンロード (461K) -
Tadashi SHIMO-OKA, Yoshio WATANABE1981 年 90 巻 5 号 p. 1297-1307
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーActomyosin Mg2+-ATPase activity was stimulated by a brain microtubule-asso-ciated protein (MAP) fraction. The stimulating activity of the MAP fraction was abolished by boiling and trypsin treatment, suggesting the presence of a protein factor. The factor stimulated actomyosin Mg2+-ATPase activity stoichiometrically by about four times in the optimum conditions (50-75 mM KCl, pH 6.6). The stimulating factor was coprecipitable with actomyosin and was found to be a pair of high-molecular-weight polypeptides (mol wts, 240, 000 and 235, 000). The polypeptides were not associated with microtubules or myosin, but with fibrous actin. In column chromatographies used for purifying the stimulating factor, the amount of polypeptides coincided with the stimulating activity. Increases in both specific activity and the amount of the paired polypeptides were nearly parallel in the process of the purification. A purified fraction (65% pure with respect to the paired polypeptides) showed a 56-fold increase of the specific stimulating activity as compared with the initial brain supernatant. The two peptides were similar but not identical with filamin and spectrin in terms of electrophoretic mobility. Hence, the pair of polypeptides was identified as an actin-binding protein newly
found in brain.抄録全体を表示PDF形式でダウンロード (4030K) -
Yasuharu TAKADA, Kunio HIWADA, Tatsuo KOKUBU1981 年 90 巻 5 号 p. 1309-1319
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーAngiotensin converting enzyme [EC 3. 4. 15. 1] was solubilized from the membrane fraction of human kidney cortex using trypsin and purified to homogeneity by DEAE-cellulose, hydroxylapatite and DEAE-Sephadex A-50 column chromatographies, preparative isoelectric focusing, and Sephadex G-200 gel filtration. The final recovery of the enzyme was 13.9%. The molecular weight of the enzyme was estimated to be 199, 000 by a sedimentation equilibrium method. A value of 170, 000 was obtained for the reduced and denatured enzyme by dodecylsulfate-polyacrylamide gel electrophoresis. The enzyme was a glycoprotein consisting of a single polypeptide chain with an isoelectric point of 5.10. Neutral sugar accounted for 13 % per weight of the enzyme.
The purified enzyme had a specific activity of 96.0 μmolimin/mg protein for hippurylhistidylleucine. The Km value, Kcat value and hydrolytic coefficient (Kcat/ Km) of the enzyme for hippurylhistidylleucine were 2.0mM, 545 s-1 and 273mM-1•s-1, respectively.
Rabbit antibody against the human kidney converting enzyme inhibited the activities of the enzymes from human lung and serum as equally as that from human kidney, but not those from sheep, dog, or rat sera. The human kidney and lung converting enzymes were immunologically identical on double immunodiffusion analysis.抄録全体を表示PDF形式でダウンロード (2191K) -
II. Kinetic Studies with a Reaction System Containing Physiological Concentrations of LigandsKatsura IZUI, Masaaki TAGUCHI, Minoru MORIKAWA, Hirohiko KATSUKI1981 年 90 巻 5 号 p. 1321-1331
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーIn an attempt to clarify the kinetic properties of Escherichia coli phosphoenolpyruvate (PEP) carboxylase [EC 4. 1. 1. 31] in vivo and to evaluate the physiological significance of the individual effectors, saturation curves were obtained for each ligand with reaction mixtures (pH 7.3) containing “physiological concentrations” of the other ligands in various combinations. As the “physiological concentrations” of ligands, which are defined as the concentrations of ligands found in the glucosegrown cells, the following values were employed: PEP, 0.2mM; acetyl-CoA(CoASAc), 0.4 mM; fructose 1, 6-bisphosphate (Fru-1, 6-P2), 2.0mM; GTP, 1.0mM; L-aspartate, 1.0mM; L-malate, 1.0mM (Morikawa, M., Izui, K., Taguchi, M., & Katsuki, H. (1980) J. Biochem. 87, 441 149).
In the absence of any activator the enzyme activity was very low. CoASAc was the most powerful activator. The other two activators (Fru-1, 6-P2 and GTP) exhibited essentially no activation alone, but produced a strong synergistic activation with CoASAc. The severe inhibition by L-aspartate or L-malate was effectively alleviated only through this synergistic action of the activators. The presence of all three activators decreased the half-saturation concentration (S0.5) of PEP from 15mM to 0.35mM and increased the maximal velocity attainable at infinite concentration of PEP about 15-fold. In the system containing all five effectors, which is close to the in vivo condition, the saturation curve of PEP was sigmoidal with a Hill coefficient of 1.6 and with an S0.5 value of 3.0mM, which is about 15-fold larger than its “physiological concentration.” On the basis of the rate-concentration curve for each effector obtained with the reaction mixture containing PEP and the other effectors at “physiological concentrations, ” it was suggested that all five effectors significantly contribute to the enzyme activity in vivo. Palmitoleate, another activator of the enzyme, showed no activation in such a reaction mixture.
The sensitivity of the enzyme to the “physiological concentration” of each effector was also observed in an in situ system using permeabilized E. coli cells, where the enzyme concentration was as high as in vivo.抄録全体を表示PDF形式でダウンロード (843K) -
Hiroko TAKAHASHI, Yoichi HANAOKA1981 年 90 巻 5 号 p. 1333-1340
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーFour gonadotropin components were purified from the basic protein fraction of bullfrog pituitary glands. All the components had high potencies not only in the ovulation assay using Xenopus laevis ovaries but also in the competitive binding assay for rat follicle-stimulating hormone (FSH) using Xenopus laevis testis. The isoelectric points of the four components determined by isoelectric focusing were at pH 8.8, 9.0, 9.1, and 9.3, respectively. No appreciable difference in the physical and chemical properties other than isoelectric points was found among the four components. Their Stokes radius estimated by gel filtration was 2.36 nm. This value was appreciably lower than that reported for mammalian luteinizing hormone (LH). On sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, reduced preparations of the four components each showed two bands, indicating a subunit structure similar to that of mammalian gonadotropins. Their mobilities were greater than those of mammalian LH. The molecular weights of subunits estimated by high-speed gel filtration in 6M guanidine hydrochloride were lower than those of mammalian LH. The amino acid composition of the four basic components of bullfrog gonadotropin was appreciably different from that of mammalian LH.抄録全体を表示PDF形式でダウンロード (1951K) -
Toshihiro TSUDZUKI1981 年 90 巻 5 号 p. 1341-1349
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーIt was examined whether or not enzymically synthesized L-dopamine could be immediately incorporated and stored in synaptic vesicles within a synaptosomal cytoplasm fraction isolated from the rat brain. The synaptosomal cytoplasm was isotonically prepared by disrupting the synaptosomes in 0.32M sucrose using an N2-gas pressure homogenizer. Synaptic vesicles present in the isolated synaptosomal cytoplasm appeared to be osmotically sensitive (i.e., not leaky vesicles) under iso-osmotic conditions. DOPA decarboxylase [EC 4. 1. 1. 28] present in the soluble cytosol region synthesized L-[14C]dopamine from added L-[14C]DOPA. However, the newly synthesized L-[14C]dopamine was not immediately incorporated in the synaptic vesicles within the synaptosomal cytoplasm during the reaction period of 20min at 30°C. Plain synaptic vesicles prepared by the method of Kadota and Kadota (10) also failed to take up the newly synthesized dopamine under the same conditions. It was found that soluble protein(s) in the synaptosomal cytoplasm was able to bind as much added L-[3H]dopamine as 35 pmol per mg of protein, and the binding ability seemed to inhibit the amine uptake by synaptic vesicles within the synaptosomal cytoplasm. The [3H]dopamine bound to the protein(s) became unbound in the presence of Ca2+ (5mM). This effect of Ca2+ may have an important role in the releasing process of neurotransmitters.抄録全体を表示PDF形式でダウンロード (1674K) -
Nobuyuki YAMAMOTO, Michiki KASAI1981 年 90 巻 5 号 p. 1351-1361
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe characterization of the cation channel in sarcoplasmic reticulum (SR) vesic'es was performed by measuring the choline influx. The choline influx in SR vesicles was measured by following the change in light scattering intensity using a stopped flow apparatus. From the analysis of the initial rate of choline influx, the following results were obtained. (1) The choline influx was activated by extravesicular Ca2+with an apparent dissociation constant of 8.3x10-7M and was inhibited through two steps with inhibition constants of 2.6 x 10-5M and 7.4x10-4M. (2) The dependence of choline influx on the ion concentration followed the Michaelis-Menten kinetics with a half-saturation constant of 30mM. (3) The choline influx was inhibited by lowering the pH. (4) The activation energy of choline influx was 3.5 kcal/mol. (5) The choline influx was strongly blocked by Cs+ with an inhibition constant of 10mM. These properties of the choline transporting system in SR vesicles are similar to those of the cation channel reported by Miller (J. Membrane Biol. (1978) 40, 1-23). Lastly, the effect of extravesicular Ca2+ on the choline transport can be explained by an allosteric model.抄録全体を表示PDF形式でダウンロード (757K) -
Takahiko HOSHITA, Nobue HARADA, Itsuko MORITA, Kenji KIHIRA1981 年 90 巻 5 号 p. 1363-1369
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe absorption of bile alcohols by rat small intestine was studied using in vivo and in vitro preparations. In bile duct cannulated rats, intraduodenal administered bile alcohols were certainly absorbed and excreted in the bile, although the absorption was not so efficient as for bile acids. The absorption during a 24 h period for free bile alcohol, bile alcohol 3-sulfate, and bile alcohol 3-glucuronide was 60%, 31%, and 30% of the dose, respectively, compared with 98% for taurocholate. In situ recirculation experiments demonstrated the linear relationship between perfusate concentration and intestinal absorption for unconjugated and 3-sulfated bile alcohols. Everted gut sacs of rat ileum actively transported a bile alcohol possessing a sulfuric acid ester group on the side chain. However, free bile alcohol, bile alcohol 3-sulfate, and bile alcohol 3-glucuronide were not transported. These results indicate that bile alcohols having no negative charge on the side chain are absorbed by passive diffusion and not by active transport.抄録全体を表示PDF形式でダウンロード (529K) -
Taishin TAKUMA, Masayoshi KUMEGAWA1981 年 90 巻 5 号 p. 1371-1375
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe relationship between the actions of androgen and thyroid hormone in induction of trypsin-like esteroproteases in mouse submandibular gland was studied. Zymograms prepared using tosyl-L-lysine a-naphthyl ester as substrate after isoelectric focusing in polyacrylamide slab gel showed that the isozymes of trypsin-like esteroprotease induced by 5α-dihydrotestosterone and triiodothyronine were identical. The time courses of esteroprotease induction by these hormones were very similar, and the lag time was not shortened by treatment with both hormones. The doses of 5α-dihydrotestosterone and triiodothyronine for half-maximal induction were 0.5mg and 2.5μg per 100g body weight, respectively, and these values were not altered by simultaneous injection of other hormones. The binding capacity and affinity of androgen receptor for methyltrienolone, a synthetic androgen, were not affected by daily injections of triiodothyronine for 5 days.
These results suggest that androgen and thyroid hormone act independently, not competitively, and so the two hormones induce trypsin-like esteroproteases additively.抄録全体を表示PDF形式でダウンロード (1230K) -
Yukio KATO, Yoshio NOMURA, Mitsuko TSUJI, Hiroaki OHMAE, Takako NAKAZA ...1981 年 90 巻 5 号 p. 1377-1386
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーPreviously, we showed that a polypeptide isolated from fetal bovine cartilage stimulates proteoglycan synthesis, RNA synthesis, and DNA synthesis in cultured rabbit costal chondrocytes. In the present study, the effects of the cartilage-derived factor (CDF) were compared with those of multiplication-stimulating activity (MSA), which is thought to be included in somatomedins. CDF and MSA markedly increased [3H]thymidine incorporation into rabbit costal chondrocytes from 7-14 h after their additions. The incorporation of [3H]uridine was also increased within 1-2 h in the cells incubated with the peptides. These stimulations of nucleic acid syntheses were followed by cell division in serum-free culture medium. On the
other hand, the peptides enhanced the synthesis of sulfated proteoglycans within 7 h. Furthermore, the extent of stimulation of proteoglycan synthesis, as measured in terms of increased incorporation of [35S]sulfate, [3H]glucosamine, or [3H]serine into material precipitated with cetylpyridinium chloride, was consistently higher than the extent of CDF or MSA stimulation of total protein synthesis, as measured in terms of increased incorporation of [3H]serine into acid-insoluble cellular material. These findings suggest that the stimulation of proteoglycan synthesis by CDF and MSA is not directly linked to the peptide-induced stimulation of cell proliferation. The present study also showed that CDF is as effective as MSA in stimulating various metabolic indices in cultured chondrocytes at their optimal concentrations.抄録全体を表示PDF形式でダウンロード (788K) -
Yasuo OHNO, Hisao KATO, Takashi MORITA, Sadaaki IWANAGA, Katsumi TAKAD ...1981 年 90 巻 5 号 p. 1387-1395
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーProtein C is a precursor of plasma serine proteinases, and its active form inactivates specifically blood coagulation Factor V and Factor VIII. Since a specific and sensitive synthetic substrate for the activated protein C was not known, we studied its amidolytic activity toward 25 fluorogenic peptides of the type peptidyl-4-methylcoumaryl-7-amide (peptidyl MCA). The activated protein C, namely, bovine protein C activated by bovine α-thrombin, showed the highest activity toward Boc-Leu-Ser-Thr-Arg-MCA. The enzyme's Km and Kcat values for this substrate were calculated to be 3.3×10-4M and 8.4s-1, respectively. Optimum conditions for measurement of activated protein C activity were studied with this substrate. Optimum pH was 8.5. For the maximum activity at pH 8.5, concentrations of 0.1M NaCl and 1mM CaCl2 had to be maintained in the reaction mixture. The fluorogenic peptide Boc-Leu-Ser-Thr-Arg-MCA was successfully applied to a simple and accurate assay of protein C during its purification.抄録全体を表示PDF形式でダウンロード (1337K) -
Tatsuo YAGURA, Tomoko KOZU, Takeshi SENO1981 年 90 巻 5 号 p. 1397-1403
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe nature of the inhibitory effects of rifamycin derivative AF/013 (O-n-octyloxime of rifamycin SV) on DNA polymerases α and γ were studied. Lineweaver-Burk analysis of the inhibition of DNA polymerases with respect to a substrate and template-primer showed a different mode of inhibition by AF/013 for each: the inhibition of DNA polymerase γ was competitive with both dTTP and poly(rA)- oligo(dT), while that of DNA polymerase α was competitive with activated calf thymus DNA and non-competitive with dTTP. Further analysis of the competitive mode of the inhibition of DNA polymerase a, using poly(dT)-oligo(rA) as a template-primer, demonstrated that the primer molecule competed with AF/013. A change of effective divalent metal ion (Mn2+ in place of Mg2+) in the reaction mixture did not alter this competitive mode of inhibition with respect to the templateprimer. The results of experiments to obtain further insight into the mechanism of drug-enzyme interaction suggest that AF/013 binds tightly to DNA polymerase α, and inhibits the process of chain elongation with DNA polymerases α and γ.抄録全体を表示PDF形式でダウンロード (542K) -
Shuichi TSUJI, Shoichi ISHIURA, Megumi TAKAHASHI-NAKAMURA, Tetsuo KATA ...1981 年 90 巻 5 号 p. 1405-1411
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe effects of Ca2+ ions on the structure and activity of Ca2+-activated neutral proteinase (CANP) were examined by means of carboxymethylation.
CANP was inactivated by carboxymethylation of l mol of sulfhydryl group in the 80 K subunit. This sulfhydryl group was exposed to the solvent, and the buried single sulfhydryl group, which was exposed by addition of Ca2+ ions, was not essential for activity.
The carboxymethylated CANP (Cm-CANP), though inactive, was indistinguishable from native CANP with respect to conformation. The structural change of Cm-CANP induced by Ca2+ at various Ca2+ concentrations and pH values corresponded well to the activity-Ca2+ and activity-pH relationships of native CANP. It is suggested that the observed conformational changes were essential for the appearance of enzyme activity. The induced structural changes occurred only around a cysteine residue and Trp and/or Tyr residues, and no significant changes in the secondary structures were observed.抄録全体を表示PDF形式でダウンロード (484K) -
Toyozo SAKURADA, Akio NOMA1981 年 90 巻 5 号 p. 1413-1419
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリー1. The subcellular localization of monoacylglycerol lipase in rat adipocytes was studied. The enzyme activities were mainly found in the plasma membrane and in the cytosol fraction.
2. The properties of the monoacylglycerol lipase activities, from both subcellular sites, were almost identical, except for slight differences in thermostability.
3. It is conceivable that there may be two kinds of monoacylglycerol lipases in adipose tissue for the hydrolysis of the monoacylglycerols formed by the action of two kinds of triacylglycerol lipases, lipoprotein lipase and hormone-sensitive lipase.抄録全体を表示PDF形式でダウンロード (520K) -
Tsuguhiro KANEDA, Saiko KURODA, Osamu KOIWAI, Shonen YOSHIDA1981 年 90 巻 5 号 p. 1421-1427
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーTerminal deoxynucleotidyl transferase [EC 2. 7. 7. 31] has been purified 4, 365-fold from pig thymus. It was further separated into two molecular forms of 57, 000 and 45, 000 daltons by Sephadex G-100 gel-filtration. The former sedimented at 4.2S through a sucrose gradient, while the latter sedimented at 3.6S. By sodium dodecyl sulfate-polyacrylamide gel-electrophoresis, their molecular weights were estimated as 57, 000 and 42, 000 daltons, respectively. Thus, the large and small pig terminal deoxynucleotidyl transferases both consist of a single polypeptide of 57, 000 or 42, 000 daltons and have no subunit structure. These two forms were indistinguishable in antigenicity as examined by a neutralization assay with an anticalf terminal deoxynucleotidyl transferase antibody. The enzymatic properties of 42, 000-dalton terminal deoxynucleotidyl transferase from pig thymus were very similar to those of the calf enzyme, which has a two-subunit structure.抄録全体を表示PDF形式でダウンロード (1210K) -
Katsurni TODA, Ken-ichi ONO, Hiroshi OCHIAI1981 年 90 巻 5 号 p. 1429-1436
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーWe have studied the changes in glycoprotein patterns on the surface of Dictyostelium discoideum cells during development on a membrane filter and in suspension culture. Cell-surface carbohydrates were detected by a periodate- [3H] borohydride method.
The appearance and the increase of some glycoproteins during the early developmental stage on a membrane filter were induced 6 h after the onset of starvation. In suspension culture the same proteins appeared about 2 h earlier and less copies per cell were made. Some glycoproteins present in growth-phase cells, especially with high molecular weights, partially disappear during development on a membrane filter. However, the latter changes were not observed in suspension culture.
Pulses of cAMP, which generally accelerate cell development, induced only the appearance of one glycoprotein.
We conclude that the changes in patterns of plasma membrane glycoproteins during early development are regulated by several factors; starvation, pulses of cAMP, cell-cell contact and transfer onto a membrane filter.抄録全体を表示PDF形式でダウンロード (3885K) -
Tsutomu KODAKI, Hiroko MURAKAMI, Masaaki TAGUCHI, Katsura IZUI, Hirohi ...1981 年 90 巻 5 号 p. 1437-1444
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーA large amount of pyruvate was excreted into the medium by CP78 (rel+) cells grown on succinate when they were starved for amino acids. In contrast, no such excretion was observed with CP79 (rel-) cells. This phenomenon was also seen with two other isogenic pairs of strains: NF161 (rel+) and NF162 (rel-), and 10B601 (rel+) and 10B602 (rel-). Besides succinate, L-malate, and fumarate were effective carbon sources for the excretion, but glucose, glycerol, and acetate were not. When of-lactate was used, not only CP78 but also CP79 cells excreted pyruvate. Experiments using [1, 4-14C]succinate as a carbon source revealed that pyruvate was formed by decarboxylation of one carboxyl group of succinate and that the pyruvate excretion amounted to about 40% of the total succinate degraded. Experiments designed to elucidate the mechanism of the excretion yielded the following observations. (i) The concentration of pyruvate in CP78 cells grown on the C4-dicarboxylic acids mentioned above was not significantly changed upon amino acid starvation. (ii) Guanosine 5'-diphosphate-3'-diphosphate exerted no effect on the activities of several enzymes thought to be involved in pyruvate-related metabolism. It is suggested firstly that the excretion was not due to some impairment in the biosynthetic pathway of a particular amino acid, but was due to the stringent control of central amphibolic metabolism, and secondly that no de novo protein synthesis was involved in the excretion.抄録全体を表示PDF形式でダウンロード (1479K) -
Hiroshi SAKAMOTO, Yoshiyuki ICHIKAWA, Toshio YAMANO, Toshio TAKAGI1981 年 90 巻 5 号 p. 1445-1452
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーCircular dichroic spectra of NADPH-adrenodoxin reductase, adrenodoxin and the complex between oxidized adrenodoxin reductase and oxidized adrenodoxin were studied.
A 1:1 complex formation between adrenodoxin reductase and adrenodoxin resulted in the appearance of a distinct difference CD spectrum, which was obtained by subtraction of the sum of their CD spectra from the CD spectrum of the mixture of the two proteins. This difference CD spectrum showed a broad trough between 500 and 600 nm and CD extrema at 452, 383, and 350 nm. This spectrum may reflect a change in the environment of either or both of the protein chromophores.
By applying the curve-fitting method to analyze the ellipticity change at 452 nm of the difference CD spectrum obtained on titration of a fixed amount of adrenodoxin reductase with various amounts of adrenodoxin, we directly obtained a value of 4.56 ×10-7M (S. D. 9.58×10-8M) as the dissociation constant of the oxidized complex.
We also studied the change of the difference ellipticity induced by the addition of salt and obtained dissociation constants for the oxidized complex at various concentrations of NaCI. These values were larger than those obtained previously by kinetic methods, measurements of NADPH-cytochrome c reductase and NADPHDCIP reductase activities. Further, we examined the relationship between the change of these activities on the addition of salt and the complex concentration calculated using the dissociation constant obtained in this study and found that the change of these activities could not be attributed simply to dissociation of the oxidized complex.抄録全体を表示PDF形式でダウンロード (1567K) -
Isao CHINEN, Tuyoshi NAKAMURA, Nobuhiro FUKUDA1981 年 90 巻 5 号 p. 1453-1461
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe immature sugar cane stalks studied contained less than 7% sucrose, and showed the activities of enzymes such as invertase, α-galactosidase, α-mannosidase, β-N-acetylglucosaminidase, β-glucosidase, β-xylosidase, and β-galactosidase. The α-galactosidase was highly purified by ammonium sulfate fractionation, gel filtration on a Sephadex G-100 column, ionexchange chromatography on DEAE-cellulose, and CM-cellulose columns, and heat treatment (60°C, 15min) in the presence of 0.2M D-galactose. In polyacrylamide gel electrophoresis, the purified enzyme was homogeneous, having a molecular weight of approximately 46, 000. In gelfiltration, it was approximately 47, 000. The activity was optimum at pH 4.5 and at 60°C. The purified enzyme hydrolyzed p-nitrophenyl-α-n-galactopyranoside (Km, 0.83mM; Vmax, 25.0 μmol/mg/min), raffinose (Km, 25.9mM; Vmax, 15.4 μmol/mg/min), and stachyose (Km, 13.0mM; Vmax 2.7 μmol/mg/min), in addition to melibiose, guar gum, and locust bean gum. The hydrolysis of p-nitrophenyl-α-D-galactopyranoside was markedly inhibited by HgCl2, AgNO3, p-chloromercuribenzoate (PCMB), L-ascorbic acid, melibiose, stachyose, and n-galactose. Also the purified enzyme showed a lectin activity with trypsinized erythrocytes.抄録全体を表示PDF形式でダウンロード (1356K) -
Hiroshi YANAGAWA, Yoko OGAWA, Atsuo MIYAKAWA, Fujio EGAMI1981 年 90 巻 5 号 p. 1463-1478
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーDNases A1 and A2 have been purified to homogeneity from the hepatopancreas of Achatina fulica by a series of steps: acetate buffer extraction, ammonium sulfate precipitation and column chromatography on hydroxylapatite, phosphocellulose, Blue-Sepharose, and poly(A)-Sepharose. The purified enzymes are free of acidic phosphomonoesterase, phosphodiesterase, and RNase activities. They are slightly acidic glycoproteins with identical isoelectric point (6.90).
On 0.1% SDS gel electrophoresis, DNase A2 had a molecular weight of 30, 000 when dissolved in 1 % SDS, but it had molecular weights of 17, 500, 8, 000, and 4, 800 when dissolved in 1% SDS and 1% 2-mercaptoethanol. This was evidence that the enzyme consists of three different subunits joined by interchain disulfide bonds. DNases A1 and A2 are endonucleases working at acidic pH (3.5-6.0) and do not require divalent cations for their activities. The enzymes degrade poly(dA) 5 times faster and poly(dT) 3 times faster than heat-denatured DNA under optimal conditions but do not appreciably digest poly(dG) and poly(dC). We developed an analytical procedure for oligodeoxynucleotides by high-performance liquid chromatography. The phosphomonoester end group and the mode of degradation were examined by the method. The termini produced by the enzymes have 3'-phosphoryl and 5'-hydroxy end groups. The products of exhaustive hydrolysis contain di-, tri-, tetra-, and pentanucleotides and mononucleotide was barely detected. The hydrolyzing activities of DNases A1 and A2 are stimulated by polyamines such as spermine, spermidine, and putrescine, but are inhibited by synthetic polynucleotides and various drugs. Adenosine deaminase highly active on oligoadenylic acids was found in a crude DNase A fraction. The enzyme preparation has higher activity on 3'-adenylic acid than on 5'-adenylic acid. The first adenosine residue of oligoadenylic acids was deaminated considerably more rapidly than the second or succeeding ones.抄録全体を表示PDF形式でダウンロード (1917K) -
Hiroshi YANAGAWA, Yoko OGAWA, Fujio EGAMI1981 年 90 巻 5 号 p. 1479-1485
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe degradation of heat-denatured and native calf thymus DNAs, poly(dA-dT), poly(dA-dC)•poly(dG-dT), and poly(dG-dC) by DNase A has been investigated with the main aim of providing background information for studying the specificity of the enzyme. The specificity of the DNase A was studied by determining the base compositions of 5'- and 3'-terminal nucleotides of oligonucleotides released by the enzyme. The 5'- and 3'-terminal nucleotide compositions were found to vary in the average chain length (_??_n) range of 45 to 7 for degradations of heatdenatured calf thymus DNA at pH 4.0, 4.5, 5.0, and 5.5. When the heat-denatured DNA was digested at the optimal pH (pH 5.0), at the _??_n=45 dAdo was predominant (39%) and dThd was minor (9%) in 5'-terminals, whereas dThd was predominant (43%) and dAdo was minor (9%) in 3'-terminals. At _??_n=7, dAdo was even more predominant (49%) and there was very little dThd (7%) in 5'-terminals. No preference was seen in 3'-terminals. This finding indicates that change in the specificity takes place during digestion. The compositions of 5'- and 3'-terminal nucleotides of oligonucleotides released from poly (dA-dT) by exhaustive digestion of the enzyme showed pronounced preferences for dAdo in 5'-terminals (81%) and dThd in 3'-terminals (78%).
The hexamer or above deoxyadenylate and the nonamer or above of thymidylate were good substrates of the enzyme.
It can be concluded that DNase A is a novel cluster-specific DNase which recognizes and cleaves sequences of oligodeoxyadenylate and/or sequences of oligothymidylate.抄録全体を表示PDF形式でダウンロード (520K) -
Yasuhide SEKI, Naoko SOGAWA, Makoto ISHIMOTO1981 年 90 巻 5 号 p. 1487-1492
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーSiroheme extracted by acetone/HC1 treatment of sulfite reductase from yeast and purified by column chromatography catalyzed the reduction of sulfite to thiosulfate and sulfide when coupled with a hydrogen-hydrogenase-methyl viologen system. The activity increased with decrease in pH from 7 to 4, and an apparent Km value of 50mM for sulfite was obtained. In contrast to sirohydrochlorin plus Fe2+, addition of inorganic iron or 2, 2'-bipyridine prior to the reduction reaction had scarcely any effect on the sulfite-reducing activity of siroheme. Hydroxylamine was reduced by siroheme at a much faster rate than sulfite, and the rate increased with increase in pH from 6 to 9. Siroheme extracted from Chromatium vinosum strain D sulfite reductase also reduced sulfite to thiosulfate and sulfide.抄録全体を表示PDF形式でダウンロード (386K) -
Hiroko TODA, Michio YAZAWA, Kiyoshi KONDO, Tamotsu HONMA, Kozo NARITA, ...1981 年 90 巻 5 号 p. 1493-1505
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe complete amino acid sequence of scallop calmodulin was determined by isolating and sequencing the peptides obtained after cyanogen bromide cleavage and tryptic digestion. The protein consisted of 148 amino acid residues and its amino(N)-terminus was blocked with an acetyl group. Scallop calmodulin lacked tryptophan and cysteine residues and contained one mol each of N_??_-trimethyllysine (Tml) and histidine residues per mol of the protein. Scallop calmodulin contained only one tyrosine residue, while vertebrate calmodulins contain two. On comparing its amino acid sequence with that of bovine calmodulin, three amino acid substitutions were found at positions 99 (Tyr→Phe), 143(Gln→Thr), and 147(Ala→Ser).抄録全体を表示PDF形式でダウンロード (878K) -
Masafumi TSUJIMOTO, Keizo INOUE, Shoshichi NOJIMA1981 年 90 巻 5 号 p. 1507-1514
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーInteraction of CRP with liposomes was studied by agglutination, complement consumption, and the binding assay.
Liposomes composed of phosphatidylcholine and stearylamine were agglutinated with CRP. The interaction of CRP and liposomes was further demonstrated by the consumption of hemolytic complement activity. Stearylamine could be substituted by cetyltrimethylammonium bromide, indicating that positively charged molecules were required for the interaction.
Agglutination of liposomes caused by CRP was dependent on the fatty acid composition of phosphatidylcholine, cholesterol content and temperature. Differential scanning calorimetry measurement revealed that both the agglutination of liposomes and the consumption of complement caused by CRP required the fluid state of membranes.
Particle electrophoresis measurement showed that stearylamine in fluid state membranes was more dispersed than in rigid state membranes indicating that lateral distribution of stearylamine affected the reactivity of liposomes. The mode of complement activation by the interaction of CRP with positively charged liposomes was similar to that observed in the system of protamine and heparin.抄録全体を表示PDF形式でダウンロード (576K) -
Yasuzo NISHINA, Kiyoshi SHIGA, Hiromasa TOJO, Retsu MIURA, Hiroshi WAT ...1981 年 90 巻 5 号 p. 1515-1520
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe resonance Raman (RR) spectra of the complexes of D-amino acid oxidase (DAO) with benzoate derivatives were measured. The RR spectra of complexes of DAO with benzoate derivatives excited at 514.5 nm are similar to one another and also similar to that of oxidized flavin. In the cases of DAO-o-NH2-benzoate and DAO-o-OH-benzoate complexes, however, the line at 568 or 565cm-1, derived from the benzoate derivative, was intensified. In the case of DAO-o-NH2-benzoate complex, which has an intense charge-transfer absorption band, the resonance enhancement of the Raman lines at 1583 and 568cm-1 in the RR spectrum excited at 632.8 nm is striking. The former line is known to involve the vibrational displacements of the N (5) and C (4a) atoms of isoalloxazine and the latter is considered to be derived from a ring deformation mode of o-NH2-benzoate. This suggests that the o-NH2-benzoate molecule lies along the N (5)-C (4a) bond and parallel to the flavin face. A Raman line derived from o-OH-benzoate in the RR spectrum of DAO-o-OH-benzoate complex excited at 514.5 nm was detected. This result supports the view that the complex has a charge-transfer band, as has been pointed out by Massey and Ganther.
Also, the spectrum of quasi-DAO-o-OH-benzoate complex is identical with that of the complex of DAO, suggesting that the active sites of these two enzymes have similar structures.抄録全体を表示PDF形式でダウンロード (443K) -
Miho TAKAHASHI, Mariko KOBAYASHI, Tsuneko UCHIDA1981 年 90 巻 5 号 p. 1521-1527
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーPurified nuclease TT1 from Thermus thermophilus HB8 has multimolecular weight forms, each of which is composed of three different subunits, α (10.8×104), β (7.8×104), and γ (4.1×104). The molecular weights of this enzyme were estimated by gel filtration, polyacrylamide gel electrophoresis and equilibrium sedimentation. It was found that most of the enzyme has a molecular weight of about 22×104 being a monomer having the subunit composition of αβγ. The remaining part of the enzyme has larger molecular weights and is considered to be size-isomers of αβγ. The α-helical content, 5.5-6.5%, and the β-structure, about 28%, were estimated from the CD spectrum at 4°C.抄録全体を表示PDF形式でダウンロード (1089K) -
Mutsumi SUGITA, Tokuko YAMAMOTO, Shigeki MASUDA, Osamu ITASAKA, Taro H ...1981 年 90 巻 5 号 p. 1529-1535
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe spermatozoa of the fresh-water bivalve, Hyriopsis schlegelii, had a complex spectrum of neutral glycosphingolipids. CDS, CTS, and CTeS were isolated as minor components (together they accounted for 3% of the total neutral glycolipid fraction), while CMS and the glycolipids (Lipid I, II, and III) with larger saccharide chains that were previously characterized (1-4) occurred as major ones. The chemical structures were determined by employing exoglycosidases, partial acid hydrolysis and permethylation analysis. The proposed structures are as follows: Manα(1-3)Manβ(1-4)Glc-Cer for CTS, and Manα(1-3)[Xylβ(1-2)]Manβ(1-4)Glc-Cer for CTeS. CTeS was the first example of a tetraglycosyl ceramide containing xylose. In another fraction, two types of diglycosyl ceramides were obtained as a mixture, and identified as Manβ(1-4)Glc-Cer and Galβ(1-4)Glc-Cer, respectively.
The long-chain base was predominantly composed of sphingosine, and the acyl groups were mainly of palmitate and stearate.抄録全体を表示PDF形式でダウンロード (2435K) -
Shigeyuki FUKUI, Hiroaki YOSHIDA, Ikuo YAMASHINA1981 年 90 巻 5 号 p. 1537-1540
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe sulfohydrolytic activity to degrade active sulfate (3'-phosphoadenosine 5'- phosphosulfate, PAPS) and its precursor, APS (adenosine 5'-phosphosulfate), with a pH optimum at 9.5 was found to be widely distributed in various tissues of rats. In the liver, the activity was located in plasma membranes and endoplasmic reticula. Triton X-100 solubilized rough and smooth endoplasmic reticula gave two peaks of the activity on gel filtration, both of which had nucleotide pyrophosphatase activities, hydrolyzing the pyrophosphate linkages of ATP, NAD, and UDP-Glc, and the phosphodiester linkage of PNTP (p-nitrophenyl-thymidine 5'-monophosphate) besides PAPS and APS.抄録全体を表示PDF形式でダウンロード (323K) -
Akemi SUZUKI, Tamio YAMAKAWA1981 年 90 巻 5 号 p. 1541-1544
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーOur previous studies demonstrated clearly the presence of asialo GM1 in the intestinal mucosa and microvillus membranes of mouse. The present work demonstrated the distribution of asialo GM1 as well as the entirely complementary distribution of Forssman antigen in the small intestine of mouse by immunofluorescence staining. Clear staining of the brush border and basolatcral membranes of epithelial cells, the cell membranes of cryptic cells and also some secretory granules was observed with the purified rabbit anti-asialo GM1 IgG and fluorescence-conjugated goat antirabbit IgG. On the other hand, Forssman antigen was demonstrated neither on the brush border membranes nor cryptic membranes, but demonstrated distinctly in the mesenchymal tissue. Such a remarkable difference of distribution of the two glycolipids, which are biosynthesized by different pathways from a common precursor glycolipid (lactosylceramide), indicates that the expression of sugar transferases for elongation of carbohydrate units may be regulated by precise genetic information during organ differentiation.抄録全体を表示PDF形式でダウンロード (995K) -
Kuniji TANAKA, Hirohisa KAWAGUCHI, Kyoko OHNO, Keiko SHOHJI1981 年 90 巻 5 号 p. 1545-1548
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーA novel enzyme for producing difructose anhydride IV (di-D-fructofuranose 2, 6':6, 2' dianhydride) from levan of Bacillus mesentericus was prepared from a culture of Arthrobacter ureafaciens, and the conditions and method to determine the enzymatic activity were determined. Levan of Bacillus subtilis also yielded the difructose dianhydride, but inulin, inulobiose, sucrose, levanbiose, and levantriose did not produce it on enzymic reaction under similar test conditions.抄録全体を表示PDF形式でダウンロード (277K) -
Yoshiaki TAKAHASHI, Kikuo OGATA1981 年 90 巻 5 号 p. 1549-1552
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーAfter rat liver polysomes were irradiated with UV light at 254 nm for 2 h, a crosslinked poly(A)-containing mRNA-protein complex (mRNP) was prepared and protein moiety was labeled with125I. After RNase treatment, its protein moiety was analyzed by two-dimensional polyacrylamide gel electrophoresis followed by radioautography. There were radioactive spots which extended from the positions of those of S3/S3a, S6, L5, and L6/L7 towards the origin. In the case of UV irradiation for 30min, radioactive spots extending similarly from those of S3/S3a, and L5 were observed. Radioactive areas on the two-dimensional gel in the case of irradiation for 2 h were further analyzed by SDS polyacrylamide gel electrophoresis. The peaks of radioactivity were detected at the protein band containing L6, that containing S3a and L5 and that containing S6, L7, and L8. It was proposed that S3a, S6, L5, and L6 proteins, according to the proposed uniform nomenclature (McConkey et al. (1979) Mol. Gen. Genet. 169, 1-6 (l)), were cross-linked to mRNA by UV irradiation.抄録全体を表示PDF形式でダウンロード (1033K) -
Noboru OTOTANI, Zensaku YOSIZAWA1981 年 90 巻 5 号 p. 1553-1556
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーPorcine intestinal heparin was partially digested with a purified heparinase and an octasaccharide with high-affinity for antithrombin III was isolated from the digest by gel filtration, followed by affinity chromatography on a column of Sepharose 4B coupled with antithrombin III. The anticoagulant activity determined by the activated partial thromboplastin time method of the octasaccharide was 240 units/mg. Fifty percent inactivation activities of the octasaccharide for thrombin and factor Xa in the presence of antithrombin III were 2 and 6.5 times, respectively, higher than those of the initial heparin. These data indicate that the octasaccharide possesses the critical structural integrities required for manifesting these biological activities.抄録全体を表示PDF形式でダウンロード (270K) -
Shoichi ISHIURA, Kazunori HANADA, Masaharu TAMAI, Keiko KASHIWAGI, Hid ...1981 年 90 巻 5 号 p. 1557-1560
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーThe role of intracellular calcium-dependent proteinase(s) has been investigated in intact rat muscle. When calcium ions were introduced into intact muscle in vitro with ionophore A23187, Z-line loss and concomitant release of α-actinin into the medium were observed. The calcium-induced release of α-actinin was not diminished in the muscle with in vivo-injection of a thiol protease inhibitor, E-64-c. Intramuscular concentrations of E-64-c were also measured after pulse labeling with [3H]E-64-c followed by subcellular fractionation. Most of the inhibitor was localized in the cytosol, not in the lysosome.
Therefore, we conclude that cytosolic as well as lysosomal proteinases in muscle are not inhibited by the in vivo labeling of the protease inhibitor (10mg/kg).抄録全体を表示PDF形式でダウンロード (325K) -
Akira MURASUGI, Chiaki WADA, Yukimasa HAYASHI1981 年 90 巻 5 号 p. 1561-1564
発行日: 1981/11/01
公開日: 2008/11/18
ジャーナル フリーWhen S. pombe is cultured in a medium containing a high concentration of CdCl2 (1mM), it grows for over 20 h accumulating Cd2+ in the cells. Simultaneously, Cd-binding peptides (Cd-BP1 and -BP2) are synthesized, and accumulated depending on the time after addition of Cd2+ to the culture medium. Apparent molecular weights of Cd-BP1 and -BP2 are 4, 000 and 1, 800, respectively. Both Cd-BPs are composed of common unit peptides, confirmed by chemical and physicochemical analyses.抄録全体を表示PDF形式でダウンロード (296K) -
1981 年 90 巻 5 号 p. 1565a
発行日: 1981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 (35K) -
1981 年 90 巻 5 号 p. 1565b
発行日: 1981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 (35K)
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