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Volume 65 (1969)
- Issue 6 Pages 849-
- Issue 5 Pages 667-
- Issue 4 Pages 497-
- Issue 3 Pages 329-
- Issue 2 Pages 159-
- Issue 1 Pages 1-
Volume 65, Issue 6
Displaying 1-18 of 18 articles from this issue
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ISAMU SHIIO, RYUICHI MIYAJIMA1969 Volume 65 Issue 6 Pages 849-859
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAspartate kinase [ATP: L-aspartate 4-phosphotransferase, EC 2. 7. 2. 4] has been partially purified about 10 fold from sonic extracts of Brevibacterium fiavum. This enzyme requires ATP and a divalent cation for its activity and SO4 ion stimulates the reaction significantly.
While L-lysine or L-threonine at 1mM gave only 10-20% inhibition, simultaneous addition of the two amino acids at 1mM each produced over 90% inhibition. L-Isoleu-cine or L-valine had an activating effect and recovered the activity from the concerted inhibition caused by L-lysine Plus L-threonine.
From kinetic analysis, the aspartate kinase reaction was explained as a Michaelis type rapid equilibrium system with random addition of the two substrates (L-aspartate and ATP). The inhibition by L-lysine was competitive to L-aspartate, of mixed type to ATP, while the inhibition by L-threonine was partially competitive to both L-aspar-tate and ATP. The concerted inhibition was explained by an increased affinity of one inhibitor to the enzyme which was caused by binding of another inhibitor. Thus, when L-threonine had been bound to the enzyme or the enzyme-ATP complex, L-lysine showed approximately 17 and 100 times higher affinity to these enzyme complexes, respectively.
The activation by L-isoleucine was K type. The relationship between L-isoleucine and L-lysine was competitive. The reversal of the concerted inhibition by L-isoleucine was explained by a competitive relationship between L-isoleucine and L-lysine.View full abstractDownload PDF (736K) -
MASASHI SATO, HIKARU OZAWA1969 Volume 65 Issue 6 Pages 861-867
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDetermination of ubiquinone was carried out in mitochondrial fractions of two parasitic nematodes, lung worms, Metastrongylus elongatus, and round worms, Ascaris lumbricoides var. suis.
In mitochondria from the lung worm, which lives under aerobic conditions, ubi-quinone-9 was detected, whereas in mitochondria from the round worm, which lives under anaerobic conditions, ubiquinone was not detected. The concentration of ubiquinone-9 in the lung worm was 0.72μmole/g of mitochondrial protein. In place of ubiquinone, a quinone analogous to rhodoquinone was detected in the round worm.
The present experiment has demonstrated the occurrence of ubiquinone in the phylum Nemathelminthes, and furthermore has provided the first evidence showing the occurrence of a rhodoquinone analogue in the animal tissue.View full abstractDownload PDF (444K) -
II. A Possible Conformational Change Induced by ATPTATSUHISA YAMASHITA, SHIZUKO KOBAYASHI, TAKAMITSU SEKINE1969 Volume 65 Issue 6 Pages 869-877
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSMyosin was treated with p-nitroaniline diazonium fluoroborate (NDF) to modify its structure chemically.
1. NDF at a molar ratio of 3.6:1 to myosin subunit strongly inhibited the Ca++-ATPase [EC 3. 6. 1. 3] activity, but slightly suppressed the EDTA-ATPase activity. Conversely, when 1mM ATP or ADP was present in the coupling reaction mixture, the diazonium compound suppressed the activity of EDTA-ATPase but only slightly inhibited Ca++-ATPase. Two moles of ATP per mole of myosin subunit were enough to bring the above-mentioned change of the inhibition pattern of ATPase to completion. AMP had no effect.
2. With 1mM ITP or PPi in the presence of Mg++, NDF inactivated both Ca++-and EDTA-ATPases.
3. Inhibition of the ATPase activity by NDF decreased with increasing pH of the coupling reaction mixture.
4. The catalytic properties of myosin modified with NDF in the presence and absence of ATP also differed in pH dependence, the Km values and Mg++- and K+-ATPase activities. The properties of the former were almost the same as those of myosin modified with diazobenzene-p-sulfonate which had been investigated by us.
On the basis of the experimental data obtained, the conformational change induced by ATP and its analogues was discussed.View full abstractDownload PDF (584K) -
AKIYUKI OHKUBO1969 Volume 65 Issue 6 Pages 879-888
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSHuman mercaptalbumin was treated with glutathione or cysteine at pH 7.0 to yield nonmercaptalbumin monomer in which the sulfhydryl group was bound with glutathione or cysteine, respectively. The ultraviolet difference spectra and optical rotatory disper-sion of human serum albumin monomer, human serum albumin dimer, and human nonmercaptalbumin monomer, i.e., human mercaptalbumin-glutathione or human mer-captalbumin-cysteine compound were studied and compared.
It was proved that: A) One tryptophanyl residue was exposed on the surface of protein and two tyrosyl residues, one of which was exposed and the other half-buried, were located near the sulfhydryl group in native human serum albumin. B) These chromophores in the neighborhood of the sulfhydryl group were inverted in the interior of the molecule with a fairly large decrease in the specific optical rotation of human serum albumin at 233mμ as the sulfhydryl group was bound with a thiol group (from -9, 300° to -8, 000° by combination with glutathione, to -7, 400° with cysteine, and to -8, 700° with human serum albumin monomer).
It seems to be possible to conclude that: 1) The reactive sulfhydryl group in the native albumin is located at the border between a polar helical segment and a hydrophobic area on the surface of the protein. 2) The half-buried tyrosyl residue near the sulfhydryl group is accommodated in a hydrophobic area located in the neigh-borhood of a helical segment or between helical segments of the protein fold, and may be the tyrosyl residues in the albumin molecule which is located at the closest position to the tryptophanyl residue.View full abstractDownload PDF (744K) -
IV. The Role of CoAHIDEO INOUE, TAKUO TSUNEMI, FUJIO SUZUKI, YOSHIRO TAKEDA1969 Volume 65 Issue 6 Pages 889-900
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe details of the reaction mechanism of ATP citrate lyase [EC 4. 1. 3. 8], especially on the mode of action of CoA, were investigated using a single homogeneous prepara-tion from rat liver. The results obtained are: (1) The enzyme rapidly catalyzes the acetyl-CoA-CoA exchange, but not the acetyl-CoA-acetate exchange. (2) The com-bination of 14C-acetate to the protein occurs on incubation of 14C-acetyl-CoA with the enzyme, showing the formation of an enzyme-acetate complex. When this complex is isolated by gel filtration and then incubated with oxaloacetate, 14C-citrate is formed. The formation of an enzyme-oxaloacetate complex is not detected. (3) The enzyme easily combines with free CoA or CoA that is released from acetyl-CoA during the for-mation of the enzyme-acetate complex. This binding of CoA to the enzyme does not depend on the presence of added ATP, and the bound CoA is freely exchangeable with exogenous CoA. These data indicate that the linkage between enzyme and CoA is not energy-rich, unlike that between enzyme and acetate.
From these findings, it is concluded that enzyme_??_, enzyme_??_and enzyme_??_are involved as intermediates in the reaction, and the possibility of an involvement of citryl-CoA as a reaction intermediate is excluded.
An additional role of CoA, besides that of an acetyl acciptor, is also discussed.View full abstractDownload PDF (932K) -
I. Isolation and Purification of Active SubstancesHIROYUKI SATO, TAKENORI TANIMURA, ZENZO TAMURA1969 Volume 65 Issue 6 Pages 901-906
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Three antibacterial substances A, B and C were found in the cultured broth of Streptomyces carzinostaticus var. F-41. The main activity resided in substance A.
2. Substance A was isolated and purified in the yield of 40 to 50% from the original culture fluid. The homogeneity of the purified A was demonstrated by electrophoresis, ultracentrifugation and chromatography on the column of DEAE-cellulose, Sephadex G-75, Hydroxyapatite and Amberlite CG-120.View full abstractDownload PDF (2523K) -
MASACHIKA IRIE1969 Volume 65 Issue 6 Pages 907-913
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Ribonuclease from Aspergillus saitoi [RNase M, EC 2. 7. 7. 17] was found to be inhibited competitively by various nucleosides. Among four common ribonucleosides, the inhibitory effect was in the following order, A>C>G>U.
2. When the base was the same, deoxyribonucleoside was found to be less effective than ribonucleoside. It was suggested that the RNase M bound with 2'-OH group of ribonucleoside in some way.
3. α-Anomers of adenosine and guanosine were less effective than the natural β-anomers. This indicates that the RNase M requires the nucleoside the proper orienta-tion of the sugar 2'-OH or 3'-OH and base moiety.
4. L-β-Cytidine-5'-phosphate was less effective than D-β-cytidine-5'-phosphate (CMP). From this result, it was suggested that the RNase M requires nucleosides the proper spacial relationship among base, 2'-OH and ether oxygen.
5. When mixed with nucleosides, the RNase M gave similar difference spectra to those observed by mixing with nucleotides.
6. pKi-pH profiles of adenosine, cytidine and guanosine were studied and it was found that the bends concaved side upwards at pH 6.0 observed in the case of pKm- or pKi-pH profile of nucleotide were not observed.
7. Inactivation of the RNase M by iodoacetate was protected by the presence of adenosine, but not by photooxidation.
8. From the results obtained here, possible mode of binding of nucleosides with the RNase M was discussed.View full abstractDownload PDF (481K) -
AKIRA BABA1969 Volume 65 Issue 6 Pages 915-923
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSFerritin was crystallized in a form of hexahedron from coelomic fluid of a shellfish (Corbicula sandai) living in fresh-water. In the purification and crystallization process, ammonium sulfate was used exclusively. The crystalline preparation contained 4.5% of iron and 0.6% of phosphorus. Absorption spectrum of this preparation in the ultra-violet and visible regions showed a shoulder around 280mμ and no absorption band at other wavelengths. The presence of iron core or micells characteristic of ferritin was confirmed by electron microscopy. Molecular weight of this protein determined by the sedimentation equilibrium method was approximately 43.5×l04.View full abstractDownload PDF (2439K) -
YASUO KAGAWA1969 Volume 65 Issue 6 Pages 925-934
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe target molecular weights of mitochondrial ATPase [EC 3. 6. 1. 3] in the inner membrane in situ, in purified state and in reconstructed membrane were determined by means of electron irradiation with a linear accelerator. The target molecular weights of mitochondrial ATPase were irrespective of its state, close to the molecular weight of purified mitochondrial ATPase determined with other methods than the irradiation. The bead-like structure of the inner membrane of mitochondria, which may represent the ATPase molecule, was lost by the irradiation. Those activities that depend on the conservation of high energy state, such as the respiratory control ratio, the 45Ca accu-mulation and the swelling contraction, were affected by the low irradiation dose, despite their relationships to the ATPase.View full abstractDownload PDF (1808K) -
NOBUHITO SONE, EISUKE FURUYA, BUNJI HAGIHARA1969 Volume 65 Issue 6 Pages 935-943
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. By disintegration of mitochondria from Endomyces magnusii using French Press, an ATPase [EC 3. 6. 1. 3] protein was extracted in latent form. The protein was purified by fractionation with polyethyleneglycol-MgSO4 without inducing ATPase activity. The enzyme was highly purified by chromatography on a DEAE-cellulose column, and at the same time its ATPase activity was induced.
2. Activation of the latent ATPase, both in solubilized or in particle bound state, was also attained by heat treatment especially in a high concentration of salts. Mag-nesium ion was partially effective for protecting activation.
3. The properties of the yeast ATPase were compared with those of mammalian enzymes. The enzymatic properties of the former resembled those of beef heart enzyme with a few exceptions.View full abstractDownload PDF (1337K) -
MASAKO KIKUCHI, HARUHIKO NODA, KOSCAK MARUYAMA1969 Volume 65 Issue 6 Pages 945-952
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe interaction of G-actin or F-actin with H-meromyosin was studied at a low concentration of KCl (10mM) by flow birefringence, viscosity, sedimentation and electron microscopy.
The polymerization of G-actin was greatly accelerated by H-meromyosin in 10mM of KCI, and the amount of F-actin thus polymerized was approximately proportional to the amount of H-meromyosin added up to a weight ratio of 1:4.
The actin-H-meromyosin complex was found not to depolymerize even at a very low protein concentration at low ionic strength.
The number-average particle length of the actin-H-meromyosin complex was about 0.8μ and the weight-average length was about 1.6μ, when G-actin was added to H-meromyosin.
The actin-H-meromyosin complex at low ionic strength showed two schlieren peaks with about 80S and 150-200S in the sedimentation pattern. Upon addition of ATP, the peak of F-actin with 40S was observed.View full abstractDownload PDF (1173K) -
TAKAO MATSUO, ZIRO SUZUOKI1969 Volume 65 Issue 6 Pages 953-960
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe urinary excretion of 4-methylthiazole-5-acetic acid (MTA) was investigated on rabbits, dog, human subjects and rats receiving orally thiamine or thiamine tetrahydro-furfuryl disulfide (TTFD). The qualitative and quantitative determinations of MTA were performed by either direct or invert isotope dilution method following chromato-graphic separation using Amberlite CG-50 ion exchanger. In rabbits MTA accounted for 14 to 57% of radioactivity excreted into the 24hr urine and it corresponded to 4 to 35% of the administered 14C-thiamine. In a dog 6.4% of the administered thiamine was characterized as MTA in the 1-6hr urine. The occurrence of MTA was also confirmed in the human urine by the direct isotope dilution method. The average amount of MTA excreted for 3hr was 1.38mg in untreated subjects and was increased about 2-or 3-fold after ingestion of thiamine (500mg) or TTFD. From these results it is concluded that MTA is a urinary metabolite of orally administered thiamine in the mammalians so far examined.View full abstractDownload PDF (556K) -
XVII. Tyrosine, Histidine and Tryptophan Residues in Bovine Trypsin in Bovine Trypsin in the Presence and Absence of BenzamidineAYAKO MATSUSHIMA1969 Volume 65 Issue 6 Pages 961-965
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe reactivities of tryptophan, tyrosine and histidine residues in bovine trypsin [EC 3. 4. 4. 4] with a few reagents were examined in the presence and absence of benz-amidine, a strong inhibitor of this enzyme, and the following facts were revealed. Three of the total four tryptophan residues in the trypsin molecule are oxidized with H2O2-dioxane at pH 8.4 in the presence of benzamidine while all of them are oxidized with this reagent in its absence. Six of the total ten tyrosine residues are reactive with cyanuric fluoride at pH 9.1 but, with increasing the pH of the reaction mixture to 9.6 or more, an additional tyrosine residue becomes reactive in the absence of inhibitor, accompanying about 50% inactivation. The presence of benzamidine at the higher pH's suppresses the reaction of this additional tyrosine residue. Diazonium-l-H-tetra-zole at pH8.8 bisazotizes all of the ten tyrosine residues and two of the three histidine residues, and the presence of benzamidine retards these coupling reactions appreciably at lower DHT concentrations. It was deduced from these data that a tyrosine residue participates in the activity of bovine trypsin, directly as a member of the amino acid residues in the active site or indirectly as a residue maintaining the active configuration of the molecule.View full abstractDownload PDF (391K) -
FUMIO MIYAZAWA, AKIKO HARA, SHIGEO IWAHARA1969 Volume 65 Issue 6 Pages 967-970
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
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HIDEO SUGITA, YOSHIKO OKUMURA, KEIKO AYAI1969 Volume 65 Issue 6 Pages 971-972
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (152K) -
TAKUO TSUNEMI, YASUSHI DAIKUHARA1969 Volume 65 Issue 6 Pages 973-975
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (175K) -
MITSURU UCHIYAMA, MASANORI YONAHA, MICHINAO MIZUGAKI1969 Volume 65 Issue 6 Pages 977-978
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
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YOSHIE ONO, SHOSHICHI NOJIMA1969 Volume 65 Issue 6 Pages 979-981
Published: June 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (211K)
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