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Volume 67 (1970)
- Issue 6 Pages 753-
- Issue 5 Pages 611-
- Issue 4 Pages 507-
- Issue 3 Pages 333-
- Issue 2 Pages 157-
- Issue 1 Pages 1-
Volume 67, Issue 3
Displaying 1-17 of 17 articles from this issue
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TAKEHIKO KAWANO, KAZUO MORIMOTO, YAHIRO UEMURA1970 Volume 67 Issue 3 Pages 333-342
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Urokinase inhibitor was purified 120 folds from human placental extract by acid treatment, ammonium sulfate precipitation, removal of water insoluble precipitate, adsorption of impurities on CM-Sephadex C-50, gel filtration and hydroxylapatite column chromatography. At the final step, the inhibitor was separated into two fractions (I and II) which were eluted with 0.1M and 0.2M phosphate buffer (pH 6.8) respectively.
2. Trypsin inhibition, enhancement of streptokinase activity, and thromboplastic activity, which were demonstrated in the crude placental extract, were not observed in the purified fraction. Inhibition against urokinase-activated plasmin remained to a negligible degree. It was also proved that the urokinase inhibitor from placenta was different from plasmin inhibitors in human plasma.
3. Urokinase inhibitor showed the electrophoretic behavior as α-globulin on cellulose acetate membrane. Being easily soluble in water, the inhibitor was considered to be pseudoglobulin.
4. By isoelectric focusing, both inhibitor fractions I and II showed one similar isoelectric point (around pH 4.8-4.9). The inhibitor having another isoelectric point (pH 5.8) was present in a sample before purification on hydroxylapatite column.
5. The inhibitor in each fraction was resolved in two sub-fractions by gel filtration for molecular size analysis. The molecular weight of inhibitor was estimated to be about 70, 000 in the first sub-fraction and about 43, 000 in the second (main) sub-fraction.
6. Urokinase inhibitor solution was stable at 40°C or a lower temperature in a pH range of 5.0-7.5, and also completely stable at 2°C for 12 months.View full abstractDownload PDF (719K) -
CHUN-CHANG CHANG1970 Volume 67 Issue 3 Pages 343-352
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. The exhaustive fluorescein thiocarbamylation of cobrotoxin through its free amino groups resulted in decrease in lethality to one-sixth that of native toxin without affecting the antigenic specificity. This suggests that the antigenic sites of cobrotoxin are different from the active site(s) of toxicity.
2. Reduced S-carboxymethylated cobrotoxin was completely devoid of lethality and antigenic activity. This indicates that the intact disulfide bonds which maintain the specific secondary structure are essential for the venom toxicity as well as antigenicity.
3. No significant differences were observed between the immunochemical properties of cobrotoxin and fluoresceinthiocarbamyl cobrotoxin as indicated by the results obtained from the precipitin reactions, immunodiffusion and immunoelectrophoresis, reactivities with Fab fragments of the purified antibody, and measurement of the antibody-binding sites in each antigen molecule. Fluoresceinthiocarbamyl cobrotoxin is a good antigen for the production of anti-cobrotoxin antibody.View full abstractDownload PDF (1594K) -
OSAMU KANAMITSU, YOICHIRO KURODA, HIUGA SAITO, YONOSUKE IKEDA1970 Volume 67 Issue 3 Pages 353-361
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSA temperature sensitive mutant was derived from Bacillus subtilis Marburg strain. The characters of this mutant were as follows:
(1) The temperature sensitive mutational site was located near the SM locus on the chromosome.
(2) When incubated at 50°C, the cells ceased growth in one hour, and lost viability thereafter.
(3) When the cells were preincubated at 50°C for 30min and thereafter incubated at 37°C, active alkaline phosphatase [EC 3. 1. 3. 1] was not formed.
(4) When 70-S ribosomes from the mutant cells were treated at 50°C for 10-20min, they were dissociated into 50 S and 30 S to a greater extent than those of parental strain, as judged from sedimentation profiles in the sucrose density gradient centrifugation.
(5) When the ribosomes were treated with ribonuclease [EC 2. 7. 7. 16] after heat treatment at 50°C for 5-10min in the presence of 2×10-4M Mg2+, more acid-soluble materials were produced than those of parental strain.
(6) When the ribosomes were treated at 45°C or 50°C for 2-5min, activity of polypeptide synthesis (incorporation of 14C-phenylalanine directed by poly U) was lost to a greater extent than those of parental strain. For example, by heating at 50°C for 2min, 75% of the activity was lost in the mutant while less than 50% was lost in the parental strain.
It was concluded from these results that the mutant isolated might produce temperature sensitive ribosomes.View full abstractDownload PDF (636K) -
MITSUMASA ISHIDA, KOZO HAMAGUCHI, TOKUJI IKENAKA1970 Volume 67 Issue 3 Pages 363-371
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe interaction of trypsin [EC 3. 4. 4. 4] with soybean trypsin inhibitor (STI)** was investigated by circular dichroism (CD)** and fluorescence techniques. The CD spectra of trypsin, STI and STI modified by cleavage of the arginyl-isoleucyl peptide bond, and the change in the CD spectrum accompanying the interaction of trypsin with STI at various pH values were studied in detail. Analysis of these CD spectra showed that the formation of modified STI is optimal at pH 4.0. This finding is in accord with that recently reported by Laskowski, Jr. and co-workers. The pH dependence of the change in the fluorescence spectrum accompanying the interaction of trypsin with STI was parallel with that of the CD spectrum.View full abstractDownload PDF (522K) -
NOBUYOSHI SHIMIZU, HIROSHI HAYASHI, KIN-ICHIRO MIURA1970 Volume 67 Issue 3 Pages 373-387
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSIn order to understand the role of a common sequence -GpTpψPpCpGp- in tRNA molecules, the inhibitory effects of the purified tetranucleotide TpTpψCpGp were studied for aninoacyl-tRNA synthesis and for the aminoacyl-tRNA binding to tRNA-ribosome complex. Since TpψpCpGp did not inhibit the aminoacylation of tRNA, it is unlikely that the TpψpCpGp sequence is involved in the functional sites of tRNA for an aminoacyl-tRNA synthetase.
The tetranucleotide TpψpCpGp inhibited aminoacyl-tRNA binding to mRNA-ribosome complex. This inhibitory effect was weaker than the terminal-pCpCpA-free tRNA fragments. The inhibitory effect by the latter fragments was a little weaker than the terminal adenosine-oxidized tRNA or the non-aminoacylated tRNA, with which the binding of the aminoacyl-tRNA was inhibited competitively. These results suggested that in addition to an anticodon sequence and a 3'-terminal sequence -pCpCpA, the sequence TpψpCpGp is required for the aminoacyl-tRNA binding to the mRNA-ribosome complex. It is further supported by the results of inhibition experiments with deaminated tRN A and with tRNA digested sequentially by phosphodiesterases [EC 3. 1. 4. 1].
Partially deaminated tRNA inhibited the binding of Phe-tRNA to the poly Uribosome complex, while it scarcely inhibited the binding of Lys-tRNA to the poly Aribosome complex. This indicates that the specificity of the nucleotide sequence in the anticodon is required for matching tRNA to mRNA.View full abstractDownload PDF (1132K) -
YOSHIAKI SEKI, TARO NAKAMURA, YOSHIRO OKAMI1970 Volume 67 Issue 3 Pages 389-396
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDuring experiments on biosynthesis of 2-nitroimidazole (an antibiotic, azomycin) by Streptomyces eurocidicus, the diazotizable amine was found to be accumulated in the cultured medium and partly in the mycelium. This amine has been purified in crystalline form from the medium and identified as 2-aminoimidazole by its chemical and physical characteristics.
The possible role of the amine as a precursor of 2-nitroimidazole is suggested by in vivo conversion of the amine to 2-nitroimidazole by washed cell-suspension. Possible biosynthetic pathway of the amine is discussed.View full abstractDownload PDF (574K) -
XXII. Effect of Cyanide on the Reactivities of Histidine and Tyrosine Residues in Ferrihemoglobin and FerrimyoglobinAKIKO TAKENAKA, OSAMU TAKENAKA, HIROO HORINISHI, KAZUO SHIBATA1970 Volume 67 Issue 3 Pages 397-402
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe reactivities of histidine and tyrosine residues in native structures of horse ferrihemoglobin and ferrimyoglobin and those in cyano derivatives and apoproteins of these hemoproteins were studied with diazonium-l-H-tetrazole (DHT) as the reagent, and the following facts were revealed. Approximately 28 out of the total 38 histidine residues and about 8 out of the total 12 tyrosine residues in the horse ferrihemoglobin molecule were bisazotized with this reagent, and addition of cyanide reduced the reactivities of some of these residues. The reduction was clearly observed at medium DHT concentrations, where the difference found in the degree of biscoupling between ferrihemoglobin and cyanoferrihemoglobin was 7.4 moles for histidine and 2.6 moles for tyrosine per mole of protein. In the case of ferrimyoglobin, 8-9 out of the total II histidine residues and both of the total two tyrosine residues were bisazotized, and addition of cyanide made one of these 8-9 histidine less reactive, but did not affect the reactivity of the two tyrosine residues. The reactivities of the histidine and tyrosine residues in apohemoglobin and in apomyoglobin were not much different from those of the residues in native hemoproteins, and cyanide did not affect their reactivities. This bears evidence that cyanide changes the reactivities through interaction with the heme group. A greater conformational change of ferrihemoglobin by the effect of cyanide as compared with that of ferrimyoglobin was deduced from these results.View full abstractDownload PDF (482K) -
TOHRU NISHIHARA, YUZO NOZU, YOSHIMI OKADA1970 Volume 67 Issue 3 Pages 403-413
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAmino acid sequence of the coat protein from RNA phage ZR (Group I) was partially determined. Oligopeptides in the tryptic digest from carboxymethylated coat protein were separated on columns of Dowex 50-X2 (soluble fraction at pH 3.1) and Sephadex G-50 (insoluble fraction at pH 3.1). The sum of the amino acids in the tryptic peptides accounts for all of the 129 amino acids present in the coat protein of ZR phage. Of the tryptic peptides, T1, T2, Tl0 and T11 were further digested by other proteolytic enzymes, and resulting smaller peptides were fractionated. Amino acid and terminal analyses were performed with each peptide and amino acid sequences of some of these peptides were determined.
The amino acid sequence of the coat protein from ZR phage is proposed in comparison with the primary structures of phage coat proteins belonging to the Group I. No difference between amino acid sequence of the coat protein from ZR phage and those from bacteriophages R17 and MS2 seemed to be present but ZR phage coat protein differs from that of f 2 bacteriophage by a single amino acid substitution.View full abstractDownload PDF (692K) -
III. Some Physicochemical Properties and Amino Acid Composition of Rhizopus chinensis Acid ProteaseDAISUKE TSURU, ATSUSHI HATTORI, HIKOJI TSUJI, JUICHIRO FUKUMOTO1970 Volume 67 Issue 3 Pages 415-420
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSSome physicochemical properties and amino acid composition of crystalline acid protease [Peptide peptidohydrolase, EC class 3. 4. 4] obtained from Rhizopus chinensis were determined. The s20, W and molecular weight of the enzyme were found by ultracentrifugal analyses to be 2.83 and 35, 000, respectively. The enzyme consisted of single polypeptide chain with alanine as the amino terminus and was composed of 324 residues of amino acids: Lys12, His0, Arg9, Asp43, Thr32, Ser26, Glu21, Pro14, Gly39, Ala23, 1/2Cys4, Val19, Met3, Ile21, Leu23, Tyr14, Phe16, Trp5 and amide-ammonia32. The optical rotatory dispersion parameters were: [α]D=-33, a0=-200, b0=0, λc=208mμ and [m']227mμ=-2, 100, suggesting the absence of α-helix structure in the molecule.View full abstractDownload PDF (461K) -
NOBUO SAKATO, KENKICHI TOMITA1970 Volume 67 Issue 3 Pages 421-435
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSIn the search for the specific chemical reactions of thyroid hormones with various biologically important compounds, it was found that 3, 5, 3', 5'-tetraiodothyroacetic acid (TETRAC) was decomposed through a transient color change (colorless→dark green→yellow) and lost its biological activities when it was treated with inorganic pyrophos-phate (PPi), ortho phosphate, arsenate and some nucleotides at pH 9, room temperature and in ambient light.
Of thyroactive compounds tested, only TETRAC and N-acetyl-thyroxine were reacting with PPi, while thyroxine and triiodothyronine were not. 3, 5, 3'-Triiodothyro-acetic acid was not only unreacting with PPi, but also inhibitory to TETRAC-PP, reaction.
By thin layer chromatography, gas chromatography and electrophoretic analyses of TETRAC-PPi reaction mixture, it was found that from 1 mole of TETRAC were formed about one iodine atom, about 0.25 mole of 3, 5-diiodo-4-hydroxyphenylacetic acid and a bulk of unknowns. No compounds of β-ring origin, such as hydroquinone or quinone derivatives, were identified yet. The reaction mechanism seems complicated, but ESR absorption spectra indicated a formation of a free radical during the TETRAC decomposition by PPi.View full abstractDownload PDF (1154K) -
NOBUO TANAKA, TOSHIO NISHIMURA, TADATOSHI KINOSHITA1970 Volume 67 Issue 3 Pages 459-463
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSProtein synthesis with endogenous mRNA and polyphenylalanine synthesis with poly U were inhibited by fusidic acid in a highly fractionated reticulocyte system. It interfered with ribosome-dependent GTPase activity of TF-II. The grade of inhibition by fusidic acid of the polypeptide synthesis and the GTPase reaction was parallel and it was comparable to what was observed in the bacterial system. The antibiotic did not significantly affect the puromycin reaction with polyphenylalanyl-tRNA or acetyl-phenylalanyl-tRNA, presumably attached to the donor site of ribosomes. However, puromycin reaction, enhanced by GTP and TF-II, was inhibited by fusidic acid. It suggested that the antibiotic selectively inhibited the translocation of peptidyl-tRNA from the acceptor site to the donor site on the reticulocyte ribosomes. The results were in accordance with those obtained with bacterial G factor, and indicated that TF-II may be a mammalian equivalent of G factor.View full abstractDownload PDF (327K) -
YUTAKA KOBAYASHI, KOICHI IWAI1970 Volume 67 Issue 3 Pages 465-472
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Calf thymus histone was chromatographed on a carboxymethylcellulose column eluted with a gradient of salt concentration in buffer containing 6M urea.
2. Whole histone was eluted in more than 4 peaks with up to 0.15M potassium chloride in 0.04-q sodium acetate-6M urea, pH 5.5. The order of elution of the histone fractions differed from those reported previously. Four histone fractions obtained by the method of Johns, arginine-rich, intermediate, slightly lysine-rich and very lysinerich, were further resolved with the present chromatographic system.
3. Further fractionation of the lysine-rich histone fraction was achieved by zone electrophoresis on a vertical column. A very lysine-rich histone, which was homogeneous electrophoretically, was obtained on a preparative scale.View full abstractDownload PDF (553K) -
TATSUO SENSHU, KOICHI IWAI1970 Volume 67 Issue 3 Pages 473-485
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. A new chromatographic method has been developed for the fractionation of calf thymus histone. The method consists of the elution of histone from a CM-cellu-lose column with a linearly decreasing gradient of ethanol in 2.6M formic acid containing 0.1M sodium formate. A unique elution order was found in that the fractions were eluted in the order of increasing lysine/arginine ratio.
2. With the whole histone, resolution of the earlier eluted arginine-rich histones is insufficient, but the later eluted slightly lysine-rich and very lysine-rich histones are clearly separated. The fractionation can be made more efficient by using as starting materials the histone fractions Fl, F2a, F2b, and F3 obtained by the method of Johns.
3. F2a is resolved into three subtractions, two of which are distinguishable by rechromatography but almost identical in their glycine-rich, arginine-rich composition and single-band disc-electrophoretic pattern. The third subfraction is an intermediate type having equimolar contents of arginine and lysine.
4. A typical slightly lysine-rich histone is obtained from F2b as a main fraction. It behaves as a single component in rechromatography, disc electrophoresis, and terminal amino acid analyses. Another subfraction similar to the intermediate-type subfraction of F2a is also found.
5. The possibility of subfractionation of F1 is also observed. The present method is least effective in further fractionation of F3.View full abstractDownload PDF (1664K) -
II. Isoelectric Point, Amino Acid Composition and Circular Dichroic SpectraYOSHIIE OKADA, KAZUO OKUNUKI1970 Volume 67 Issue 3 Pages 487-496
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. A crude larval cytochrome b-555 preparation was separated into two or three components by the method of isoelectric focusing. The two main components had much the same or the same absorption spectra, catalytic activity, molecular weight, amino acid composition, and circular dichroic spectra in the Soret region. For three components the pI values at 0-2°C were 4.24, 4.28, and 4.32, respectively. A number of explanations was suggested for the heterogeneity of larval cytochrome b-555.
2. The circular dichroic spectra of ferri- and ferrocytochrome b-555 suggested that the protein moiety of the molecule provided an asymmetrical environment to all of the chromophores in the molecule. With respect to the sign of the CD peak in the Soret region of hemoproteins containing protoheme several hypotheses were presented.
3. The amino acid composition of larval cytochrome b-555 was as follows: Lys5, His-, Arg2, Asp10, Thr3, Ser4, Glu12, Pro1, Gly4, Ala6, Val6, Met1, Ile2, Leu2, Tyr2, Phe3. The contents of tryptophan, cysteine and amide were not determined. In comparison with cytochromes b5 from various sources larval cytochrome b-555 contained the fewest amino acid residues (65 amino acid residues other than tryptophan and cysteine), lower contents (%) of histidine, and leucine, and higher contents (%) of alanine and valine.View full abstractDownload PDF (733K) -
TADAHIKO ANDO, JUN TAKAGI, TAEKO KOSAWA, YONOSUKE IKEDA1970 Volume 67 Issue 3 Pages 497-499
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (196K) -
YUZURU MIZUKAMI, MASAKI IWABUCHI1970 Volume 67 Issue 3 Pages 501-504
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (251K) -
TAKASHI YAMAMOTO, KAZUO OKUKUKI1970 Volume 67 Issue 3 Pages 505-506
Published: March 25, 1970
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (154K)
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