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Volume 76 (1974)
- Issue 6 Pages 1155-
- Issue 5 Pages 909-
- Issue 4 Pages 671-
- Issue 3 Pages 459-
- Issue 2 Pages 241-
- Issue 1 Pages 1-
Volume 76, Issue 4
Displaying 1-27 of 27 articles from this issue
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Seiki KURAMITSU, Kiyoshi IKEDA, Kozo HAMAGUCHI, Hajime FUJIO, Tsunehis ...1974 Volume 76 Issue 4 Pages 671-683
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe microscopic dissociation constants of Glu 35 and Asp 52 in hen egg-white, turkey egg-white, and human urine lysozymes [EC 3. 2. 1. 17] were determined by measuring the pH dependence of the circular dichroic band at 305nm, which is due to Trp 108 near the catalytic carboxyls, Glu 35 and Asp 52. The pk value of Glu 35 when Asp 52 is unionized (pkl, Glu) and that of Asp 52 when Glu 35 is unionized (pk1, Asp) for each of the three lysozymes were quite normal. However, the pk value of Glu 35 when Asp 52 is in a deprotonated form (pk2, Glu) and that of Asp 52 when Glu 35 is in a deprotonated form (pk2, Asp) were higher than the normal pk values of glutamyl and aspartyl residues. The macroscopic pK values of Asp 52 and Glu 35 were 3.4 and 5.9 for both hen and turkey lysozymes and 3.4 and 6.8 for human lysozyme at 25°C and 0.1 ionic strength. The high macroscopic pK value of about 6 which has been assigned to Glu 35 of hen lysozyme is not caused by its hydrophobic environment but by electrostatic interaction of ionized Glu 35 with ionized Asp 52. The pK shift of Glu 35 observed when N-acetyl-chitotriose binds to hen lysozyme may be interpreted in terms of a change in the electrostatic interaction between Glu 35 and Asp 52 accompanied by movement of the active-site cleft.View full abstractDownload PDF (974K) -
Takachika AZUMA, Kozo HAMAGUCHI, Shunsuke MIGITA1974 Volume 76 Issue 4 Pages 685-693
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. The dimerization of reduced and alkylated Bence Jones proteins was studied by gel chromatography and circular dichroic (CD) measurements. The association constants of two type λ proteins, as estimated by gel chromatography, were 105 and 2×106M-1 at pH 5.5, 25°C. The changes in the CD spectrum in the aromatic absorption region with protein concentration and those with pH on the alkaline side indicate that the monomeric and dimeric forms of type λ proteins are different from each other as regards the environment of tyrosyl residues.
2. The conformation of IgG molecules reconstituted from the alkylated H chain of a myeloma protein and various specimens of alkylated Bence Jones proteins was studied by CD measurements. The CD spectra of the recombinants were different depending on the specimens of L chains used, even if the antigenic type was the same. This indicates that the conformational change observed when H and L chains recombine is mainly associated with the conformational change of the variable portion of the L chain.View full abstractDownload PDF (613K) -
Shunji NATORI, Kazuhisa SEKIMIZU, Den'ichi MIZUNO1974 Volume 76 Issue 4 Pages 695-702
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSFactors stimulating RNA polymerase II [EC 2. 7. 7. 6] of Ehrlich ascites tumor cells were isolated from homogenates of mouse liver, kidney, and brain. The elution profiles of these factors from phosphocellulose were characteristic for each organ. The molecular size of RNA synthesized in the presence of these factors was much larger than that of RNA synthesized by RNA polymerase II alone.
These stimulatory factors lost some of their activity when they were preincubated with alkaline phosphatase [EC 3. 1. 3. 1], suggesting that their phosphate residues are responsible for stimulation of RNA synthesis in vitro.View full abstractDownload PDF (590K) -
VII. Double-labeling of Catalase by 14C-Leucine and 3H-δ-Aminolevulinic AcidTokuhiko HIGASHI, Fumiko KAWAMATA, Terufumi SAKAMOTO1974 Volume 76 Issue 4 Pages 703-708
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSTo investigate the mechanism of incorporation of heme into newly-synthesized catalase [EC 1. 11. 1. 6] rat liver catalase was labeled simultaneously with 14C-leucine and 3H-δ-aminolevulinic acid both in vivo and in vivo. The results obtained were as follows:
1. The kinetics of incorporation of 3H of injected δ-aminolevulinic acid into catalase on ribosomes were the same as those of incorporation of 14C-leucine.
2. In a cell-free system newly-synthesized catalase was found to be labeled with both isotopes.
3. The ratio of 3H/14C incorporated in vivo was lowest in catalase of microsomes and ribosomes, highest in that of peroxisomes, and intermediate in catalase of the soluble fraction of liver cells. The former remained almost constant, while the latter two increased to a plateau after injection of the isotopic tracers.
Results suggested that an intermediate is formed which contains less heme than in the completed molecule. Further addition of heme to this intermediate seems to occur in the cytosol, and probably also in the peroxisomes.View full abstractDownload PDF (461K) -
I. Isolation and Properties of Two Inhibitors of Kallikrein, Trypsin, Plasmin, and α-Chymotrypsin from the Venom of Russell's viper (Vipera russelli)Hidenobu TAKAHASHI, Sadaaki IWANAGA, Tomoji SUZUKI1974 Volume 76 Issue 4 Pages 709-719
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSSeveral substances which inhibit the activities of bovine plasma kallikrein [EC 3. 4. 21. 8], trypsin [EC 3. 4. 21. 4], plasmin [EC 3. 4. 21. 7], and α-chymotrypsin [EC 3. 4. 21. 1], were separated from crude venom of Russell's viper (Vipera russelli). All those inhibitors were polypeptidic substances having a molecular weight of about 6, 000 to 7, 000 and two of them, designated as inhibitors I and II, were purified extensively and their properties examined. The final preparations appeared pure on disc polyacrylamide gel electrophoresis with and without sodium dodecylsulfate (SDS). Their molecular weights were estimated by SDS-gel electrophoresis to be about 7, 200 (±1, 000). Inhibitors I and II contained 52 and 60 amino acid residues, respectively. The amino-terminal residue of inhibitor II was a single histidine and the carboxyl-terminus was lysine. The N-terminal of inhibitor I did not react with phenylisothiocyanate, suggesting that it was in a masked form. The carboxylterminus of inhibitor I was lysine.
Both inhibitors inactivated bovine trypsin, probably by formation an enzymeinhibitor complex in a molar ratio of 1:1. The K1 values of inhibitor II, measured using synthetic ester substrates, were 7.6×10-10M for bovine trypsin, 1.4×10-10M for bovine α-chymotrypsin, 2.9×10-10M for bovine plasma kallikrein, and 1.0×10-9M for bovine plasmin. Inhibitor II has no effect on the activities of thrombin [EC 3. 4. 21. 5], thiol proteinases (bromelain [EC 3. 4. 22. 4], papain [EC 3. 4. 22. 2], and ficin [EC 3. 4. 22. 3]), metal proteinases (thermolysin [EC 3. 4. 24. 4]) or exopeptidases (carboxypeptidases A and B [EC 3. 4. 12. 2 and 3]). Thus, its inhibition spectrum was very similar to that of pancreatic basic trypsin inhibitor (Kunitz type).View full abstractDownload PDF (769K) -
II. Chemical Structure of Inhibitor II Isolated from the Venom of Russell's viper (Vipera russelli)Hidenobu TAKAHASHI, Sadaaki IWANAGA, Tsunehiro KITAGAWA, Yasuji HOKAMA ...1974 Volume 76 Issue 4 Pages 721-733
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe complete amino acid sequence of proteinase inhibitor II isolated from the venom of Russell's viper was established by Edman degradation and standard enzymatic and chemical techniques. The inhibitor consisted of 60 amino acid with histidine and lysine at the NH2- and COOH-termini, respectively. It has a molecular weight of 6, 700 and was very basic. It contained 6 half-cystines in disulfide linkages; the locations of these half-cystine residues were determined by analyzing thermolytic fragments obtained from the native inhibitor II.
The overall primary structure of Russell's viper venom proteinase inhibitor II was very similar to that of bovine pancreatic basic trypsin inhibitor (Kunitz type) and cow colostrum trypsin inhibitor, having about 50 percent sequence homology. The 6 half-cystines of these three inhibitors were in the same positions in the amino acid sequences of the polypeptides. Moreover, the structural portions, which are found in the interior and the reactive site and tend to stabilize the unique structure of the Kunitz-type inhibitor, were found with extremely high sequence homology in proteinase inhibitor II. The evolution of these proteins is intriguing, since the evolutionary histories of cows and snakes are quite different.View full abstractDownload PDF (928K) -
Yukinobu HINO, Yoshio IMAI, Ryo SATO1974 Volume 76 Issue 4 Pages 735-744
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe induction by phenobarbital (PB) of microsomal drug-metabolizing enzymes in the residual portion of rat liver after partial hepatectomy was studied. Administration of PB 1hr after surgery caused only a slight increase in the microsomal content of cytochrome P-450 during subsequent 48 hr, although the cytochrome was normally induced by PB given 1 hr after sham operation. This greatly reduced responsiveness to PB immediately after partial hepatectomy was, however, restored gradually as liver regeneration proceeded, and the liver regained the full capacity of PB-induced accumulation of cytochrome P-450 by the 4th day after the operation. Microsomal NADPH-cytochrome c reductase was not inducible by PB immediately after the surgical operation in both sham operated and partially hepatectomized rats; however, it became thereafter inducible gradually, though only to slight extents. Cytochrome b5 was not inducible throughout the regenerative process. The change in the ethyl isocyanide difference spectrum of reduced cytochrome P-450 following PB injection 4 days after partial hepatectomy was essentially similar to that in PB-treated sham operated rats. But in the induction immediately after partial hepatectomy no change was observed in the difference spectrum. The specific activity of aminopyrine N-demethylase of all the microsomal preparations examined in this study was virtually proportional to the specific content of cytochrome P-450.View full abstractDownload PDF (786K) -
Taiji IMOTO, Masanori FUJIMOTO, Kazuyoshi YAGISHITA1974 Volume 76 Issue 4 Pages 745-753
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSTrp-62 oxidized lysozyme was purified by affinity chromatography. The enzymic activities of Trp-62 oxidized lysozyme toward glycol chitin and bacterial cells were examined and the Vmax values were found to be 8±1 and 29±4% of those of native lysozyme [EC 3. 2. 1. 17], respectively.
Not only substrate binding but also the catalytic importance of Trp-62 in lysozyme action was discussed. A cooperative effect of Trp-62 and lysyl amino group (s) in lytic action can explain the higher Vmax value in lysis.View full abstractDownload PDF (648K) -
Binding of Adenosine Di- and Triphosphates to Myosin during the Hydrolysis of Adenosine TriphosphateAkio INOUE, Yuji TONOMURA1974 Volume 76 Issue 4 Pages 755-764
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe amounts of ATP and ADP bound to myosin during the ATPase reaction [EC 3. 6. 1. 3] were determined in the presence of 2mM MgCl2 and 50mM Tris-HCl at pH 7.8, using pyruvate kinase [EC 2. 7. 1. 40] and phosphoenolpyruvate to regenerate ATP and 3H-labelled ATP as the substrate. The amounts of ADP and total nucleotides bound to myosin were measured, respectively, by thin layer chromatography after stopping the reaction with TCA and by a rappid-flow dialysis method. The following results were obtained; they were all consistent with our original reaction mechanism for myosin-ATPase but in conflict with the oversimplified variant proposed by Taylor et al.
1. Binding of ATP to myosin was only observed when the amount of ATP added was more than about 0.6 mole/mole of myosin. In the presence of both 0.1 and 0.5M KCl, the maximum amount of ATP bound was about 1 mole/mole of myosin, and the dissociation constant of binding (1μM in 0.5M KCl at 0°C) was equal to the Km value of myosin-ATPase in the steady state at high concentrations of ATP.
2. When a sufficient amount of ATP was added to myosin, 1 mole of ADP bound rapidly to 1 mole of myosin during the initial phase of the reaction. Then, the amount of bound ADP decreased to the steady state level within a few minutes.
3. The amount of bound ADP in the steady state increased almost linearly with increase in the amount of ATP added, and reached a constant value when the molar concentration of ATP added was higher than that of myosin. The maximum amount of bound ADP was 1 mole/mole of myosin at KCl concentrations above 1M, and decreased with decrease in the KCl concentration. For example, in 0.125M KCl at 20°C it was 0.4 mole/mole of myosin. The amount of bound ADP decreased slightly when the temperature was raised from 0 to 30°C.
4. The rate of the ATPase reaction in the steady state was not proportional to the amount of ADP bound to myosin.View full abstractDownload PDF (750K) -
Teruyoshi ARAI, Eiji ICHISHIMA1974 Volume 76 Issue 4 Pages 765-769
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAcid carboxypeptidase [EC 3. 4. 12. 1] isolated from the culture filtrate of Aspergillus saitoi has been investigated for its use in the carboxy-terminal sequence determination of native or reduced S-carboxymethyl-lysozyme and reduced S-carboxymethylribonuclease at pH 2.5.
The sequence of the first four carboxy-terminal residues of denatured lysozyme, leucine, arginine, S-carboxymethyl-cysteine, and glycine, could be deduced unequivocally from a time release plot of an incubation mixture with the enzyme. The enzyme catalyzed the release of a small amount of leucine from native lysozyme in citrate buffer, pH 2.2, containing a small amount of the surface active agent Brij-35.
The first five residues, valine, serine, alanine, aspartic acid, and phenylalanine, were liberated rapidly from the carboxy terminus of reduced S-carboxymethyl-ribonuclease. A small amount of proline, which occupies position eight from the carboxy terminus, can be detected only after prolonged incubation for 20.5 hr.View full abstractDownload PDF (411K) -
Koichi SUZUKI, Kazutomo IMAHORI1974 Volume 76 Issue 4 Pages 771-782
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Phosphoglycerate kinase [EC 2. 7. 2. 3] was isolated in a homogeneous form from thermophilic bacteria, Bacillus stearothermophilus, and some properties were examined.
2. The enzyme was composed of a single polypeptide chain of molecular weight 42, 000. The thermophilic enzyme had many properties in common with enzymes from mesophilic sources, e. g., pH optimum, kinetic properties, physical structure, amino acid composition, etc., though it was more stable against heat and urea denaturation.
3. Analyses of the secondary structure of the enzyme showed that it was composed of 45% β-structure, 20% α-helix, and 35% random coil.
4. Titration of SH groups indicated that there was a single SH group in the molecule. The SH group reacted with various SH reagents only in the presence of denaturing reagent and did not seem to participate in the catalytic function.
5. A classification of PGK into two classes is proposed based on the number and reactivity of SH groups.View full abstractDownload PDF (849K) -
Seiichi TAKASAKI, Akira KOBATA1974 Volume 76 Issue 4 Pages 783-789
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSA very sensitive method for the quantitative determination of individual neutral and amino sugars and N-acetylneuraminic acid in complex saccharides has been developed. The usefulness of this method was confirmed by applying it to the analysis of several milk oligosaccharides of known structure.View full abstractDownload PDF (554K) -
Yoshio INOUYE, Shizuo HANDA, Toshiaki OSAWA1974 Volume 76 Issue 4 Pages 791-799
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe lipid metabolism of human lymphocytes after stimulation with Phaseolus vulgaris hemagglutinin (PHA) for 4 days was investigated by means of [6-3H]glucose and [1-3H]galactose incorporation. In the case of [6-3H]glucose incorporation, most of the radioactivity (70-80%) is incorporated into neutral lipids, especially into triglycerides. However, the incorporation of radioactivity into polar lipids, namely phospholipids and sphingoglycolipids, is somewhat enhanced in PHA-stimulated lymphocytes compared with normal lymphocytes. In contrast, only 35-40% of the incorporated radioactivity was attributable to neutral lipids in the case of [1-3H]galactose incorporation and more than 40% of the incorporated radioactivity was found in sphingoglycolipids. Among the sphingoglycolipids, the incorporation into ceramide trihexoside was most accelerated, with the following order of incorporation into other sphingoglycolipids: glucosyl ceramide > ceramide dihexoside > globoside I. Significant incorporation of radioactivity into galactosyl ceramide was not observed.View full abstractDownload PDF (637K) -
Masao KAWAKITA, Ken-ichi ARAI, Yoshito KAZIRO1974 Volume 76 Issue 4 Pages 801-809
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSEF-Tu•GTP, but not EF-Tu•GDP, was shown to interact with uncomplexed ribosomes since only the former was protected by ribosomes against inactivation by N-ethylmaleimide. However, the GTP moiety of EF-Tu•GTP was hardly hydrolyzed, indicating that the catalytic site for GTP splitting is not active in uncomplexed ribosomes.
On the other hand, the interaction of EF-Tu•GTP with ribosomal complexes carrying N-acetylphenylalanyl-tRNA or phenylalanyl-tRNA in the A site was found to lead to GTP splitting. Presumably the catalytic site for GTP splitting is activated when the A site has been occupied by appropriate tRNA ligands. However, this interaction and GTP splitting, which is uncoupled from phenylalanyl-tRNA binding, were observed only when ribosomal peptidyl transferase was inactive. On reactivation of peptidyl transferase the uncoupled GTP splitting was abolished. It is suggested that the interaction of EF-Tu•GTP with ribosomal complexes carrying either N-acetylphenylalanyl-tRNA or phenylalanyl-tRNA on the A site was eliminated in so far as their peptidyl transferase activity is retained in the active state.
From these observations it was concluded that a tight coupling of EF-Tu-dependent GTPase under physiological conditions is dependent on ligand (tRNA)-nduced activation of the catalytic site on ribosomes, and also on ligand-induced change in the affinity of ribosomes toward EF-Tu. Both are probably mediated through a ligand-induced conformational alteration of ribosomes.View full abstractDownload PDF (761K) -
I. Further Purification of High Molecular Weight Kininogen and Its Physicochemical PropertiesMasanobu KOMIYA, Hisao KATO, Tomoji SUZUKI1974 Volume 76 Issue 4 Pages 811-822
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSA method was developed for the further purification of high molecular weight (HMW) kininogen (previously named kininogen-I*) from bovine plasma. The method involved ion exchange chromatography on columns of DEAE-Sephadex A-50 and CM-Sephadex C-50 and gel chromatography on a column of Bio-Gel P-300, using hexadimethrine bromide to inhibit the conversion of prekallikrein to kallikrein [EC 3. 4. 21. 8] and diisopropylfluorophosphate to inhibit kallikrein activity. The yield was about 430mg from 12.5 liters of fresh bovine plasma. The purified HMW kininogen gave a single band on sodium dodecylsulfate-polyacrylamide gel electrophoresis, a single precipitin line on immunoelectrophoresis and a symmetrical Schlieren pattern on ultracentrifugation. However, it gave two protein bands on polyacrylamide disc gel electrophoresis at pH 8.3. On the same electrophoresis after treatment of the kininogen with purified plasma kallikrein, these two bands were replaced by a single band with faster mobility. Thus, the results suggest that these two components of HMW kininogen must be the same molecular species.
The sedimentation coefficient of purified HMW kininogen was 3.80S and its molecular weight was estimated as about 7.6×104 by sedimentation equilibrium and the Archibald method. The molecular weight estimated by gel filtration on a column of Sepharose 4B in the presence of guanidinium hydrochloride was about 8.0×104. HMW kininogen had an isoelectric point of pH 4.5, as measured by the isoelectric focusing technique. Its polypeptide content was determined to be 76% by the biuret method and 83.5% by amino acid analysis, and there was a total of 581 amino acid residues per mole of protein. HMW kininogen contained a total of 12.6% carbohydrates, consisting of hexoses (4.57%), hexosamines (3.65%), and sialic acid (4.35%). The hexosamines were glucosamine and galactosamine in a molar ratio of 1.5:1. Thus, it was concluded that HMW kininogen is a typical glycoprotein in bovine plasma.View full abstractDownload PDF (3591K) -
II. Microheterogeneities of High Molecular Weight Kininogens and Their Structural RelationshipsMasanobu KOMIYA, Hisao KATO, Tomoji SUZUKI1974 Volume 76 Issue 4 Pages 823-832
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAs described in the preceding paper, a high molecular weight (HMW) kininogen purified from fresh bovine plasma contained two components, kininogen-a and -b, which were susceptible to plasma kallikrein [EC 3. 4. 21. 8]. These components were partially separated by ion exchange chromatography on a long column of CM-Sephadex C-50. They had the same molecular weight of about 8.0×104 and their amino acid compositions and tryptic peptide maps were indistinguishable. Thus, these two kininogens seem to be essentially the same molecular species. The difference between them was found to be due to a few peptide bond cleavages along the polypeptide chain of the kininogen molecule. Evidence for this was as follows:
1. The molecular weight of kininogen-b did not change after cleavage of disulfide bridges, while that of kininogen-a clearly decreased after reduction.
2. The theoretical amount of N-terminal amino acid could not be recovered from HMW kininogens, suggesting that the N-terminal amino acid was in a masked form. However, careful analysis by the dinitrofluorobenzene method indicated the presence of 0.2 mole of serine per mole of protein. On hydrazinolysis of the HMW kininogens, leucine was identified as the C-terminal residue. Treatment of the proteins with carboxypeptidase A [EC 3. 4. 12. 2] and B [EC 3. 4. 12. 3] liberated one mole of leucine and 0.3 mole of arginine per mole of protein.
3. On treatment of HMW kininogen-a and -b with cyanogen bromide (CNBr), the former yielded kallidin in addition to a fragment from which kallidin was liberated by trypsin, while the latter yielded only the fragment containing kallidin.
On the basis of the above results, it was concluded that kininogen-b consists of a single intact polypeptide chain with a masked N-terminal and with C-terminal leucine, while kininogen-a consists of two proteins, one with a kinin moiety at the C-terminus and the other with this moiety in the interior. These microheterogeneous kininogens are thought to be derived from kininogen-b by cleavage of a few peptide bonds within polypeptide chains bridged through a disulfide linkage.View full abstractDownload PDF (767K) -
III. Structural Comparison of High Molecular Weight and Low Molecular Weight KininogensMasanobu KOMIYA, Hisao KATO, Tomoji SUZUKI1974 Volume 76 Issue 4 Pages 833-845
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. A method was developed for the purification of low molecular weight (LMW) kininogen from the same batch of bovine fresh plasma used for the isolation of high molecular weight (1IMW) kininogen.
2. Purified HMW kininogen reacted with both HMW and LMW kininogen antisera prepared, respectively, from immunized rabbit serum. Purified LMW kininogen also reacted with both antisera. Thus, the immunological properties of HMW and LMW kininogens were very similar, and their antigenic determinant groups seem to be identical.
3. On treatment of purified HMW kininogen with cyanogen bromide, a fragment was obtained containing a kinin peptide sequence, from which kallidin was liberated by trypsin [EC 3. 4. 21. 4]. The amino acid sequence of this fragment was identical to that of a fragment previously obtained from purified LMW kininogen by cyanogen bromide treatment. Moreover, tryptic peptide maps of S-carboxymethylated derivatives of HMW and LMW kininogens were very similar: the former gave 43 peptide spots and the latter 30 peptide spots; 28 peptide spots were found to be common to both kininogens. Thus, HMW kininogen seems to contain all the structural segments constituting LMW kininogen, though more detailed studies on the primary structures of the two are required to confirm this.
4. The physicochemical and chemical properties of HMW and LMW kininogens were compared and differences were discussed.View full abstractDownload PDF (1967K) -
Yoichi TAMAI, William E. M. LANDS1974 Volume 76 Issue 4 Pages 847-860
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSA microsomal preparation isolated from rat liver catalyzed the acylation of sn-glycerol 3-phosphate with both palmitoyl-CoA and oleoyl-CoA. The optimum pH was 7.5 for both acyl-CoA's, and the apparent Km values for glycerophosphate were 1.0 and 1.2mM with palmitoyl-CoA and oleoyl-CoA as acyl donors, respectively. The major reaction product was diacyl-sn-glycerol 3-phosphate in the standard assay system with either acyl-CoA.
Acyl-sn-glycerol 3-phosphate (monoacyl-GP) was added to the incubation system to trap newly synthesized radioactive intermediates. Radioactive monoacyl-GP was trapped effectively by the addition of nonlabeled 1-acyl-GP but ineffectively by adding nonlabeled 2-acyl-GP, regardless of whether palmitoyl-CoA or oleoyl-CoA was used as an acyl donor. The monoacyl-GP trapped in the presence of 1-acyl-GP was mostly the 1-acyl-GP isomer, regardless of which acyl-CoA was used. The system with added 2-acyl-GP produced various results as regards the isomeric monoacyl-GP accumulated. Although the pathway via the 2-acyl-GP could not be excluded completely in this experiment, diacyl-GP formation from glycerophosphate in rat liver microsomes appeared to occur primarily via the 1-acyl-GP as an intermediate, regardless of whether palmitoyl-CoA or oleoyl-CoA was used as an acyl donor.View full abstractDownload PDF (1181K) -
Hitoshi ARITA, Hiroshige TSUZUKI, Kazuyuki MORIHARA, Jun'ichi KAWANAMI1974 Volume 76 Issue 4 Pages 861-869
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSTwo kinds of L-arabinans have been isolated from a broth of Streptoinyces fradiae 3739 strain. Both of these arabinans, named P-1 and P-2, possess potential anti-carrageenin abscess activity. Chromatographic differences on TEAE-cellulose between P-1 and P-2 can be attributed to differences in the contents of D-galacturonic acid in these polysaccharides. From the results of various experiments, it is concluded that these L-arabinans have almost linear structures made up of 1→5 linked α-L-arabinofuranose units. D-Galacturonic acid was found to be linked to L-arabinose by an α-glycosidic linkage. The molecular weights of P-1 and P-2 were determined to be 9, 800 and 4, 700, respectively, by ultracentrifugal analyses. Periodate oxidation of these polysaccharides resulted in loss of biological activity.View full abstractDownload PDF (652K) -
Yumiko OHTA, Sachiko TANABE, Akira HACHIMORI, Tatsuya SAMEJIMA1974 Volume 76 Issue 4 Pages 871-878
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThermolysin [EC 3. 4. 24. 4] was nitrated with tetranitromethane (TNM) to elucidate the contribution of tyrosyl residues to its thermostability and enzyme action. The nitration of 3 and 6 tyrosyl residues caused a reduction of the enzyme activity to 50% and 35%, respectively. Kinetic analysis indicated that the Km value of the nitrated enzyme for benzyloxycarbonyl-Gly-Leu-NH2 remained unchanged, whereas the maximum velocity decreased on nitration compared with that of the native enzyme. Heat treatment at 80°C for 30min revealed that nitrated thermolysin is less thermostable than the native enzyme. The α-helical conformation was found to be unaffected by nitration on the basis of ORD analysis. The nitrated thermolysin did not show fluorescence due to tryptophanyl residues, indicating that tryptophanyl residues had reacted with TNM, which caused quenching of their fluorescence. This view was supported by the spectrophotometric determination of tryptophan. It was inferred that the inactivation of the enzyme is due to the nitration of tyrosyl residues rather than tryptophanyl residues.View full abstractDownload PDF (619K) -
Yoshitaka GOTO, Hiroshi NAGANO, Ryoiti SHUKUYA1974 Volume 76 Issue 4 Pages 879-881
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (223K) -
Kunio FUJIWARA, Daisuke TSURU1974 Volume 76 Issue 4 Pages 883-886
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (260K) -
Tadashi YOSHIMOTO, Daisuke TSURU1974 Volume 76 Issue 4 Pages 887-889
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (230K) -
Bronislaw ZACHARA1974 Volume 76 Issue 4 Pages 891-895
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (273K) -
Kenji TAKAHASHI, Wen-Jong CHANG, Jin-Shung KO1974 Volume 76 Issue 4 Pages 897-899
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (182K) -
Keiji IRIYAMA, Nagao OGURA, Atusi TAKAMIYA1974 Volume 76 Issue 4 Pages 901-904
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSA convenient laboratory method for partial purification of chlorophylls using dioxane is described. Chlorophyll in crude extracts with 90%-methanol or 80%-acetone was mixed with dioxane and then precipitated by drop-wise addition of water. Chlorophyll is precipitated in spherocrystalline form, leaving the bulk of carotenoids in solution. This is an efficient method for preparing chlorophyll from various plant materials, including higher plants and algae as well as photosynthetic bacteria (bac to riot hlo ro phyll).View full abstractDownload PDF (1265K) -
Hitoshi KANETA, Daisaburo FUJIMOTO1974 Volume 76 Issue 4 Pages 905-907
Published: October 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSA previous study has shown that histone deacetylase extracted from calf thymus with 0.05M phosphate buffer, pH 7.0, can deacetylate only free histone and cannot deacetylate chromatin-bound histone. An enzyme capable of deacetylating chromatinbound histone has now been extracted from calf thymus nuclei with 0.2M phosphate buffer, pH 7.0. The enzyme differs from the previously known enzyme in chromatographic behavior.View full abstractDownload PDF (237K)
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