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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 3
Displaying 1-24 of 24 articles from this issue
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Nobuyuki ITOH, Toshisuke KAWASAKI, Ikuo YAMASHINA1974 Volume 76 Issue 3 Pages 459-466
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSGlycopeptides were prepared by pronase digestion from rat liver mitochondria, and were fractionated into neutral and acidic fractions. Neutral glycopeptides accounted for 20-30% of the total glycopeptides on a molar basis, and their carbohydrate moieties consisted mainly of glucosamine and mannose. Acidic glycopeptides consisted mainly of glucosamine, sialic acid, mannose, galactose, and fucose. These mitochondrial glycopeptides were different from the corresponding glycopeptides from hepatic microsomes in size and chemical composition.View full abstractDownload PDF (609K) -
III. Isolation and Characterization of the Active-site Peptides from Bovine RenninWen-Jong CHANG, Kenji TAKAHASHI1974 Volume 76 Issue 3 Pages 467-474
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Bovine rennin [EC 3. 4. 23. 4] was inactivated by stoichiometric reaction with, 14C-labeled diazoacetylnorleucine methyl ester (DAN) in the presence of cupric ions The inactivated enzyme was hydrolyzed with pepsin [EC 3. 4. 23. 1] and the digest was fractionated by column chromatography on Sephadex G-25 and high voltage paper electrophoresis. A single labeled peptide fraction thus obtained was treated with aqueous pyridine to cleave the ester linkage between the peptide and the reagent moiety and then subjected to high voltage paper electrophoresis under the same conditions as used in the first electrophoresis. A major peptide which is thought to be the DAN reactive-site peptide, was isolated and sequenced to be Asp-Thr-Gly-Thr-Ser-Leu.
2. Bovine rennin was also inactivated by reaction with 1, 2-epoxy-3-(p-nitrophenoxy)-propane (EPNP) with incorporation of 2 EPNP moieties per molecule. The inactivated enzyme was hydrolyzed with pepsin and the EPNP reactive-site peptides were isolated in the same way as in the case of the DAN-reactive peptide. One of the peptides thus obtained was sequenced to be Asp-Thr-Gly-Ser-Ser.
3. These results show that bovine rennin has two distinct aspartic acid residues in the active site and that the β-carboxyl group of one of these residues reacts specifically with DAN and that of the other with EPNP. The amino acid sequences around these aspartic acid residues in the isolated peptides are identical with the amino acid sequences around the DAN- and EPNP-reactive aspartic acid residues in pepsin. These results thus demonstrate a close homology between pepsin and rennin.View full abstractDownload PDF (592K) -
Toshio SHIBAOKA, Keiichi MIYANO, Takehiko WATANABE1974 Volume 76 Issue 3 Pages 475-479
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Rate parameters (Michaelis constant Km and molecular activity k0) for the hydrolyses of various maltodextrins with degrees of polymerization ranging from 3 to 117 catalyzed by saccharifying α-amylase from Bacillus subtilis [EC 3. 2. 1. 1]* were determined at pH 5.4 and 25°C.
2. The values of Km (expressed in molar substrate concentration units) decreased with increasing degree of polymerization (n) of the substrate. A sharp increase was observed in the value of k0 between n=3 and 4, and it became almost constant for n greater than 4.
3. On the basis of an assumption that the breakdown rate constant of productive ES complexes is identical irrespective of substrate n and of productive binding modes (1), the n-dependence of Km and k0 leads to the conclusion that the active site of this enzyme is composed of four subsites.View full abstractDownload PDF (362K) -
Hiroaki FURUTA, Akira HACHIMORI, Yumiko OHTA, Tatsuya SAMEJIMA1974 Volume 76 Issue 3 Pages 481-491
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAcetylation of catalase [EC 1. 11. 1. 6] was carried out with both N-acetylimidazole (AI) and acetic anhydride (AA). The maximal degree of acetylation of catalase was 54% with Al and 74% with AA according to the ninhydrin method. The sedimentation coefficient (s20, w) of 54% acetylated catalase was 9.3S, which indicated only a considerable unfolding of the molecule. It showed 65% activity and about 20% reduction in α-helical conformation as compared with the native enzyme, through the conformation in the vicinity of heme groups seemed to be unchanged. Acetylation with AA to 74% caused the protein to split into two components of 9.7 and 3.6S. Each component could be isolated by sedimentation in a sucrose density gradient from 5 to 25%. The molecular weights of the two components thus obtained were estimated to be 220, 000±30, 000 for 9.7S and 61, 000±1, 000 for 3.6S by the sedimentation equilibrium method. The former was almost the same as that of native catalase and the latter was a one-fourth sized subunit. The results indicated that acetylation with AA caused partial dissociation of the catalase molecule into 1/4-sized subunits which still retained some α-helical conformation. Fluorescence analyses of tryptophanyl residues and 8-anilinonaphthalene-1-sulfonate bound to catalase indicated that the surface of the protein may be covered with hydrophilic amino acid residues. When the molecule was split into subunits by acetylation or denaturation with acid or base, the fluorescence intensity increased remarkably, suggesting that hydrophobic region (s) buried inside the molecule was exposed to the surface.View full abstractDownload PDF (850K) -
III. N-Terminal Analysis and Degradation in a Slightly Alkaline SolutionTakuji SASAKI, Haruhiko NODA1974 Volume 76 Issue 3 Pages 493-502
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe N-terminal amino acids of fibroin directly extracted from the silk gland of Bombyx mori larva were examined by dinitrophenylation and dansylation. Quantitative analysis of the N-terminal amino acids revealed that Asp and Ser were liberated to the extent of one mole each per mole of fibroin. Glycine and alanine, which were dominant in fibroin and were previously reported as the N-terminal residues, were hardly detected. Further analysis showed that aspartate and serine were derived from the large component (C-I), which was connected with the small component (E-I) by disulfide bonds to form the fibroin molecule. The latter component gave no N-terminal residue. Fibroin was degraded in a slightly alkaline solution and reinvestigated by N-terminal analysis, gel electrophoresis, and sedimentation analysis. DFP inhibited this degradation and the presence of some proteolytic enzyme was indicated.View full abstractDownload PDF (4072K) -
Limited Dependence on Molecular WeightKoei HAMANA, Koichi IWAI1974 Volume 76 Issue 3 Pages 503-512
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSChromatography on polyacrylamide gel (various Bio-Gels) in HCl, guanidine hydrochloride (GuCl), urea, or sodium dodecyl sulfate (SDS) solution and electrophoresis on polyacrylamide gel in acetic acid, acetic acid-urea, or SDS solution were examined for the separation of disulfide bond-reduced histones by molecular weight. Almost complete resolution into five or six molecular species of calf thymus, chicken erythrocyte, or Tetrahymena histone was observed on gel electrophoresis in acidic urea and alkaline SDS solutions, while gel chromatography resulted in poorer resolution. A linear relationship between the known molecular weights of calf histone species and their elution or migration behaviors (distribution or retardation coefficient) was only observed on GuCl-gel chromatography and urea-gel electrophoresis, providing a standard curve for the estimation of unknown molecular weights of chicken and protozoan histone species. On the other gels, certain histone species behaved unexpectedly, especially on gel electrophoresis in acetic acid or SDS solution, in detailed experiments. The reasons for this are discussed.View full abstractDownload PDF (2398K) -
Hyogo SINOHARA1974 Volume 76 Issue 3 Pages 513-522
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSIn dormant conidia of Aspergillus oryzae, L-leucine is transported by two distinct systems: one is inhibited by L-arginine and the other is not. The former was much more sensitive to inhibition by lithium diiodosalicylate, osmotic shock, pronase digestion, and high temperature than the latter. The activities of both systems began to increase after the conidia were incubated in phosphate buffer, in which no germination occurred. The increase in the activity of the arginine-insensitive system started soon after the incubation of the conidia, whereas the increase in that of the arginine-sensitive system was delayed for about 2 hr. No increase in these activities was observed at 0°C or in the presence of cycloheximide. When the conidia were incubated in a germination medium, essentially the same changes were observed, but to a much greater extent. It is proposed that the dormancy is maintained by a ‘membrane brake’ mechanism, and is broken by a conformational change in the membrane caused by the intake of water. The initial events of spore germination may be divided into two steps. The first step, which includes the increase in amino acid transport activities, is triggered by the intake of water and levels off after several hours unless germinants are supplied. The second step is triggered by the germinants only after the first step has proceeded to a considerable degree.View full abstractDownload PDF (681K) -
VI. Characterization of Sulfhydryl Groups in EF-Tu and EF-TsKen-ichi ARAI, Masao KAWAKITA, Shun NAKAMURA, Ikuko ISHIKAWA, Yoshito ...1974 Volume 76 Issue 3 Pages 523-534
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe properties of sulfhydryl groups in EF-Tu and EF-Ts were investigated by titra-tion with [14C] p-chloromercuribenzoate (pCMB), or 5, 5'-dithiobis (2-nitrobenzoic acid) (DTNB), and also by assessing the inhibitory effects of N-ethylmaleimide (NEM) and N-tosyl-L-phenylalanylchloromethane (TPCK) on Tu and Ts activities.
1. EF-Tu contains three sulfhydryl groups, of which two are reactive. One reactive sulfhydryl (SH1) is essential for the binding of GDP or GTP, and is protected by guanine nucleotides in the presence of Mg2+, while the other (SH2) is essential for aminoacyl-tRNA binding, and is protected by aminoacyl-tRNA only when EF-Tu exists as EF-Tu•GTP. The third sulfhydryl group in EF-Tu is non-reactive, probably being buried in the interior of the molecule, and can be titrated only after complete denaturation of the protein.
2. EF-Ts contains two sulfhydryl groups of which one is reactive, while the other is non-reactive in the native configuration of the protein. The former is essential for interaction with EF-Tu and is protected by EF-Tu.
3. Of the five sulfhydryl groups present in EF-Tu•EF-Ts, a complex between EF-Tu and EF-Ts, two are non-reactive, and may correspond to SH3 of EF-Tu and the non-reactive thiol group in EF-Ts. The other three thiols are reactive, but two of them react sluggishly ; SH1 of EF-Tu is protected by EF-Ts, and conversely, the reactive thiol in EF-Ts is protected by EF-Tu.
4. TPCK is found to react selectively with SH2 but not with SH1 when incubated with free EF-Tu in which both SH1 and SH2 are exposed.
5. Kinetic studies indicate that the reactivity of SH2 is higher in EF-Tu•GTP than in EF-Tu•GDP, when measured by titration with [14C]pCMB or by inactivation of the phenylalanyl-tRNA binding activity of Tu with NEM or TPCK. These results suggest the presence of conformational differences between EF-Tu•GTP and EF-Tu•GDP near SH2, i.e., near the site interacting with aminoacyl-tRNA.View full abstractDownload PDF (936K) -
Kunihiko IZUMI1974 Volume 76 Issue 3 Pages 535-544
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSProton NMR spectra of peracetylated methyl D-gluco-, D-galacto-, and D-mannopyranosides and their 2-acetamido-2-deoxy derivatives were measured at 100 MHz in the presence of a shift reagent, tris (2, 2-dimethyl-6, 6, 7, 7, 8, 8, 8-heptafluoro-3, 5-octanedionato)-europium. Chemical shifts and bound chemical shifts to lower fields were determined by first-order analysis for all the ring, methyl, acetyl, and amido protons in these compounds. Comparison between the neutral and the amino sugars showed that the signals of H-2, H-3, H-4, and H-6 (H-6') in the former and H-1, H-2, H-3, N-acetyl, and amido protons in the latter were more markedly shifted than others by the addition of the reagent. Differences in the bound chemical shifts were also marked between the two anomers and the greatest shifting ability was displayed by H-2 and N-acetyl protons in α-anomers of the amino sugars. From these results, the most probable coordination structures for the complexes between the sugars and the reagent were inferred, and their application for analysis of amino sugars appeared possible.View full abstractDownload PDF (601K) -
Noboru OTOTANI, Zensaku YOSIZAWA1974 Volume 76 Issue 3 Pages 545-551
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe structures of two sulfated unsaturated disaccharides, compounds 12 and 15a, isolated from eliminase [EC 4. 2. 2. 7] digests of ω-heparin (whale heparin), were determined by enzymatic and chemical methods and by proton magnetic resonance spectral analyses.
Digestion of compounds 12 and 15a with α-L-Δ4, 5-iduronidase produced N-sulfated glucosamine and N-, 6-O-disulfated glucosamine, respectively, together with sulfated a-keto acid, indicating the Δ4, 5-hexosyluronic acid moiety of these compounds to be 4-deoxy-sulfoxy-α-L-threo-hex-4-enopyranosyluronic acid.
Methyl di-O-methyl-N-, O-diacetylglucosaminides, prepared from hydrogen-reduced 12 and 15a by desulfation, N-acetylation, permethylation, methanolysis, and acetylation, in succession, were identified as α, β-methyl 3, 6-di-O-methyl-N, 4-O-diacetylglucosaminides by gas chromatography. This finding provided definite evidence for 1→4 Δ4, 5-hexosyluronic acid linkages.
In addition to the above observations, proton magnetic resonance spectral studies indicated that the structures of compounds 12 and 15a were 2-sulfoamino-2-deoxy-4-O-(4-deoxy-2-sulfoxy-α-L-threo-hex-4-enopyranosyluronic acid)-D-glucose and 2-sulfoamino-2-deoxy-6-sulfoxy-4-O-(4-deoxy-2 -sulfoxy-α-L-threo-hex-4-enopyranosyluronic acid)-D-glucose, respectively. The former compound was shown to be a novel disaccharide. The high yield of this compound suggests that it is derived from a major repeating unit of ω-heparin.View full abstractDownload PDF (482K) -
Yuzuru AKAMATSU1974 Volume 76 Issue 3 Pages 553-561
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSUnder osmotically stabilized conditions E. coli can grow with an unsaturated fatty acid content of 8 to 11% in the membrane. These membranes are fragile due to the abnormally low content of unsaturated fatty acids. Thus, the unsaturated fatty acids in the membrane do function to stabilize the membranes against osmotic pressure and to increase the efficiency of biochemical reactions in the membrane.View full abstractDownload PDF (716K) -
Purification and Enzymatic Properties of Cellulase of Fusarium monilformeKunio MATSUMOTO, Yasuo ENDO, Nobuo TAMIYA, Masatoshi KANO, Kango MIYAU ...1974 Volume 76 Issue 3 Pages 563-572
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSCellulase [EC 3. 2. 1. 4] of a phytopathogenic fungus Fusarium moniliforme was purified by ammonium sulfate fractionation and column chromatography on Amberlite CG-50, DEAE-Sephadex A-25, and hydroxyapatite. The enzyme was finally separated into two fractions (Cellulases-I and -II). Cellulase-I was purified further by hydraxyapatite column chromatography, yielding cellulase-I'. Its activity was enhanced about 10-fold over that of the starting material, using sodium carboxymethylcellulose as a substrate. Cellulase-I' was homogeneous on disc gel electrophoresis and ultracentrifugal analyses. The optimum pH for cellulase-I' activity was found to be at. 4.5, and the enzyme was stable at pH 3-8. The optimum temperature for cellulase-I'activity was found to be 60°C, and heating at 60°C for 120min caused little loss of activity. Cellulase-I' was inhibited by Hg2+, N-bromosuccinimide and sodium picryl sulfate, while it was activated slightly by Co2+, Zn2+, hydroquinone, and ascorbic acid. Ca2+, Cu2+, Ba2+, and Cd2+ had no effect on the enzyme activity. EDTA and thiol agents such as PCMB and monoiodoacetic acid also had no effect on the enzymeactivity. The molecular weight of cellulase-I' was estimated to be about 25, 000 by Sephadex G-100 gel filtration. E2801%1cm was calculated to be 15.0, and the ratio of E280/E260 of cellulase-I' was 1.74. The carbohydrate content of cellulase-I' was found to be about 26% as glucose, determined by the orcinol-sulfuric acid method. Cellulase-I' showed powerful activity toward insoluble celluloses such as Avicel and filter paper, whereas the enzyme was inactive toward cellobiose, salicine, sucrose, soluble starch, mannan, inulin, p-nitrophenyl α-D-glucopyranoside, and p-nitrophenyl β-D-glucopyranoside. Hence, the enzyme was practically free from β-glucosidase [EC 3. 2. 1. 21] activity.View full abstractDownload PDF (1944K) -
Akira OGAMO, Hiroko WATANABE, Kinzo NAGASAWA1974 Volume 76 Issue 3 Pages 573-582
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSSulfated heteropolysaccharide-peptide complexes were purified from Pronase-digested acetone-defatted powder of oyster viscera. The acidic fractions were isolated by precipitation with cetylpyridinium chloride. After gel filtration on Sephadex G-100, two sulfated fractions were finally isolated by elution from DEAE-Sephadex with sodium chloride. Each fraction exhibited a single band in cellulose acetate electrophoresis at pH 2.0 and pH 9.0.
The two fractions were rich in neutral sugars and were similar to each other in containing glucose, galactose, mannose, fucose, rhamnose, arabinose, and xylose as neutral sugar components. Sulfate was present in these fractions to the extent of 2.3 and 2.9%. The content of total hexosamine and the molar ratio of glucosamine to galactosamine in the two fractions were markedly different. Sialic acid could not be detected in either of the fractions. Total protein ranged from 2.4 to 3.9%. Aspartic acid, serine, threonine, glutamic acid, and alanine were the main constituent amino acid residues in both fractions.
Both fractions exhibited A blood group activity.View full abstractDownload PDF (3117K) -
I. Isolation and Characterization of Detergent-sensitive MutantsMitsuko ONO-ONITSUKA, Hiromi B. MARUYAMA1974 Volume 76 Issue 3 Pages 583-591
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1) Based on selection through differential sensitivity to sodium dodecylsulfate (SDS) and Triton X100, 12 ethyl methanesulfonate-induced Escherichia coli mutants with altered cell surfaces were isolated and classified into 3 groups: a) sensitive only to 5% SDS, b) sensitive to 0.5% SDS but insensitive to 5% Triton, c) sensitive to both 0.5% SDS and 5% Triton. Three representative strains of each group were characterized in comparison with the parent strain K-12: W3110 and with an oleic acidrequiring mutant, ONO-3, which also showed a slightly enhanced sensitivity to SDS.
2) The strain ONT-3 of group c) was found to be most susceptible to cell lysis caused by detergents or membrane-active antibiotics among the 5 strains tested, but had the highest resistance to a cell-wall active antibiotic, cycloserine.
3) The strain ONS-2 of group b) was second most susceptible to SDS-induced cell lysis. Bacitracin and novobiosin, which are cell-wall synthesis inhibitors also having the activity to attack plasma membrane, inhibited this strain more than any other mutants tested.
4) SDS-semisensitive strains, ONS-39 of group a), as well as ONO-3, showed surface properties almost the same as those of the parent strain, except for the sensitivity to SDS.
5) No essential difference in macromolecular synthesis or in cellular metabolic activity was found between these 4 mutants and the parent strain.View full abstractDownload PDF (635K) -
II. Induction and Subcellular Location of Lipid Peroxidation ActivityTakashi MIURA, Ryo SATO1974 Volume 76 Issue 3 Pages 593-601
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAlthough the NADPH-dependent lipid peroxidation activity of yeast homogenate is low during exponential growth under aerobic conditions, it begins to increase at the late log phase and continues to increase even after cessation of cell division. Restoration of active growth after transfer of the stationary culture to a fresh medium is accompanied by a rapid decline of the lipid peroxidation activity. When a growing culture is transferred to a medium free of carbon sources, peroxidation activity is induced within a few hours and reaches the same level as that attainable by the normal stationary culture. Addition of cycloheximide or blasticidin S to a growing culture results in complete inhibition of protein synthesis within 2 hr, but a low amino-acid incorporation activity is recovered 8 to 10 hr after the drug treatment and this activity persists for a considerable period. Under these conditions, the lipid peroxidation activity begins to increase concomitant with the restoration of aminoacid incorporation. During normal cultivation, the NADPH-cytochrome c reductase activity of cell homogenate increases in parallel with the lipid peroxidation activity, and upon cell fractionation both activities are recovered in the “microsomal” fraction.View full abstractDownload PDF (1766K) -
Dennis B. McNAMARA, Jagat N. SINGH, Naranjan S. DHALLA1974 Volume 76 Issue 3 Pages 603-609
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSBoth Ca2+ and Mg2+ stimulated ATP hydrolysis by the dog heart sarcolemmal fraction. The mean Km values were 0.90 and 0.95mM and the mean Vmax values were 17.2 and 16.0μmoles P1/mg per hr for the membrane Ca2+ ATPase [EC 3. 6. 1. 3] and Mg2+ ATPase respectively. Other divalent cations such as Mn2+, Co2+, and Ni2+ were also found to stimulate ATP hydrolysis in this fraction. Excess of ATP was inhibitory to the ATP hydrolysis due to various divalent cations. Ni2+, Co2+, Mg2+ and Mn2+ were shown to depress the ATP hydrolysis due to Ca2+. The Ca2+ ATPase activity in the heart membranes was also inhibited by Na+ whereas K+ had no effect. The adenylate cyclase activity of the heart membranes was increased by about 35% and 4 fold by epinephrine and NaF respectively. These agents increased Vmax (286 pmoles cyclic AMP/mg per min) without any changes in the Km value (0.08mM) for ATP. The activation of adenylate cyclase [EC 4. 6. 1. 1] due to epinephrine was blocked by a well known β-adrenergic blocking agent, propranolol.View full abstractDownload PDF (505K) -
Fumiyuki YAMAKURA, Yasunori IKEDA, Kinuko KIMURA, Taiji SASAKAWA1974 Volume 76 Issue 3 Pages 611-621
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. The pyruvate-aspartic β-semialdehyde condensing enzyme was purified about 1, 000-fold from cell-free extracts of Bacillus subtilis PCI-219. The molecular weight of the condensing enzyme was determined to be 124, 000±2, 000 by gel filtration on a Sephadex G-200 column.
2. The enzyme exhibited a sharp pH optimum at pH 9.5 in glycine-NaOH buffer. The Km values for pyruvate and ASA were determined to be 1.07×10-3M and 3.13×10-3M, respectively.
3. The enzymatic activity was inhibited by α, ε-diketopimelic acid and glyoxylic acid and was activated by isocitric acid. Double-reciprocal plots showed that diketopimelic acid was a competitive inhibitor with respect to ASA and pyruvate, and its Ki values were determined.
4. A comparison of the condensing enzyme from vegetative cells with that from sporulating cells was made, but no difference was observed as regards molecular weight or in the effects of diketopimelic acid, pyridine-2, 6-dicarboxylic acid (dipicolinic acid), L-lysine, and diaminopimelic acid.
5. The o-aminobenzaldehyde method for determination of 2, 3-dihydropyridine-2, 6-dicarboxylic acid (dihydrodipicolinic acid) was modified in order to give more satisfactory results for strict determination of the condensing enzyme activity.View full abstractDownload PDF (816K) -
Tetsuji SAEKI, Makoto HORI, Hamao UMEZAWA1974 Volume 76 Issue 3 Pages 623-629
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSNucleoside diphosphate kinase [EC 2. 7. 4. 6] from Escherichia coli B was partially purified, and kinetic studies of the action of desdanine were carried out. Desdanine was found to be an irreversible and specific inhibitor of the enzyme. The inhibition progressed with time and also depended on temperature. Adenosine triphosphate (ATP) but not thymidine diphosphate (dTDP) protected the enzyme from the action of desdanine. Some properties of the enzyme were also determined.View full abstractDownload PDF (500K) -
Its Role in Supplying Nucleoside Triphosphates in Cells under Anaerobic ConditionsTetsuji SAEKI, Makoto HORI, Hamao UMEZAWA1974 Volume 76 Issue 3 Pages 631-637
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSEscherichia coli cells grown under anaerobic conditions contained an increased level of pyruvate kinase [EC 2. 7. 1. 40] in contrast to a markedly decreased level of nucleoside diphosphate kinase [EC 2. 7. 4. 6]. Pyruvate kinase of E. coli, irrespective of the growth conditions of cells from which the enzyme was derived, showed a broad specificity for nucleoside diphosphates as acceptors of energy-rich phosphate from phosphoenolpyruvate. Cells growing under anaerobic conditions were less sensitive to desdanine, a specific inhibitor of nucleoside diphosphate kinase, than cells growing under aerobic conditions.
These findings strongly suggest that pyruvate kinase takes the place of nucleoside diphosphate kinase in supplying nucleoside triphosphates in E. coli under anaerobic conditions.View full abstractDownload PDF (552K) -
I. Purification and Multiple FormsKeiichi MATSUDA, Eiichi MISAKA1974 Volume 76 Issue 3 Pages 639-649
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSCathepsins A, B, C, and D were purified from the lysosome fraction of rat liver by ammonium sulfate fractionation, followed by Sephadex G-200 and DEAE-Sephadex chromatography, and evidence was presented for multiple forms of each cathepsin.
By gel filtration, each of cathepsin A, B, and D was separated into several fractions. Molecular weights estimated by the chromatography were 100, 000, 180, 000, 400, 000 for AI, AII, AIII, and 24, 000, 50, 000 for BI, BII, respectively, 150, 000 for C, and 24, 000 for the main peak of D (DI).
On DEAE-Sephadex chromatography, several peakes of cathepsin C, named CI, CII, CIII, appeared. The chromatographic pattern changed considerably in the presence of dithioerythritol, and cathepsin C became separable from cathepsin A.
By the above procedures, cathepsin A was purified 800- to 3, 000-fold, and cathepsin B, C, and D were purified 2, 100-, 480-, and 200-fold, respectively, over liver homogenate.
Cathepsin A multiple forms (isoenzymes), AI, AII, AIII appeared to have immunologically identical moieties in common, on the basis of immunodiffusion analysis. Their molecular weights suggest that they are monomer, dimer, and tetramer.
Cathepsin CI, CII, and GIII showed the same precipitin line in immunodiffusion analysis, and appear to be immunologically closely related, if not identical.View full abstractDownload PDF (2303K) -
Functional -SH Groups in Tubulin for PolymerizationRyoko KURIYAMA, Hikoichi SAKAI1974 Volume 76 Issue 3 Pages 651-654
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (258K) -
H. Hasko PARADIES1974 Volume 76 Issue 3 Pages 655-659
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSValyl-tRNA synthetase [EC 6. 1. 1. 9] from E. coli has been purified and crystallized. From still X-ray photographs, approximate cell dimensions have been obtained. The cell dimensions are a=125.0Å, b=154.0Å, and c=91.0Å with β near 90°, indicating an orthorhombic (monoclinic) unit cell. The rapid deterioration of the crystals in the X-ray beam made it impossible to determine the unit cell unequivocally. The crystalline material was proved to be valyl-tRNA synthetase by polyacrylamide gel electrophoresis.View full abstractDownload PDF (2207K) -
Akio INOUE, Toshiaki ARATA, Yuji TONOMURA1974 Volume 76 Issue 3 Pages 661-666
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (384K) -
Emi KIN, Yasuhiro ANRAKU1974 Volume 76 Issue 3 Pages 667-669
Published: September 25, 1974
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDiphenyl phosphorazidate, a new organic azide, was found to be a potent inhibitor for certain energy transformation reactions. It inhibits NaN3-sensitive, membranebound ATPase [EC 3. 6. 1. 3] of Escherichia coli and the synthesis of ATP without interfering with succinate oxidation by intact cells. It inhibits selectively the active transport reaction for isoleucine, not for proline, by intact E. coli cells.View full abstractDownload PDF (210K)
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