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Volume 82 (1977)
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Volume 82, Issue 2
Displaying 1-38 of 38 articles from this issue
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Characterization and FunctionKoscak MARUYAMA, Saburo MATSUBARA, Reiji NATORI, Yoshiaki NONOMURA, Su ...1977 Volume 82 Issue 2 Pages 317-337
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (5511K) -
Comparative BiochemistryKoscak MARUYAMA, Fumiko MURAKAMI, Kazuyo OHASHI1977 Volume 82 Issue 2 Pages 339-345
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSConnectin, an elastic protein, was isolated from both skeletal and cardiac muscles of various species of vertebrates, and also from smooth muscles (gizzard) of the chicken. The amino acid compositions of these preparations were very similar. Connectin was also obtained from claw and tail muscles of the crayfish, but preparations from clam adductor muscles and insect thoracic muscles were heavily contaminated with collagen and resilin, respectively.Connectin-like protein was obtained from cell membranes of erythrocytes and fluorescent anti-connectin staining suggested that it is located on the cytoplasmic surface of the membrane. An attempt to isolate an elastic protein from insoluble residues of amoebae of the slime mold andthose of bacterial cell body (Salmonella) was inconclusive. The present comparative bio-chemical study has shown that connectin or connectin-like protein is widely distributed in various types of muscles and in some nonmuscle cells.View full abstractDownload PDF (995K) -
Its Abundance in Cardiac MyofibrilsKoscak MARUYAMA, Sumiko KIMURA, Masaaki KURODA, Shizuo HANDA1977 Volume 82 Issue 2 Pages 347-350
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSPure myofibrils were isolated from bovine heart by sucrose layer ultracentrifugation. Cardiac myofibrils thus prepared contained more protein as insoluble stroma than skeletal muscle. The insoluble stroma largely consisted of connectin, an elastic protein of muscle. The connectin content in cardiac myofibrils was about 18% of the total myofibrillar protein and was three times that in skeletal myofibrils. In view of the role of connectin as an elastic component of muscle, the abundance of connectin in cardiac myofibrils may be responsible for keeping myofibrils short at rest. This would account for the more effective tension generation in cardiac muscle on passive stretching due to blood inflow (Stirling's law).View full abstractDownload PDF (268K) -
Kazuo HOSOI, Takao UEHA1977 Volume 82 Issue 2 Pages 351-358
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe rate of in vivo synthesis of the protein contained in the granules present in convoluted tubular cells of mouse submandibular glands (granule-protein synthesis) was measured with [3H]leucine and an antiserum specific to components contained in the granules (granulecomponents). Eight days after the castration of male mice, the rate of granule-protein syn-thesis decreased to half of that of normal male mice. But then, the rate of the castrated male mice increased to approximately twice as high as that of normal male mice 6 days after injection of testosterone propionate (0.4mg/mouse) and then decreased gradually, reaching a similar value to that of normal male mice 25 days after injection. Using extracts from the glands, the rate of protein synthesis was also measured (total protein synthesis). The ratio of rate of granule-protein synthesis to total protein synthesis decreased from 0.3 to 0.1 with castration, and with the castrated male mice, it increased to 0.45 with testosterone injection, but not with 17β-estradiol. With normal female mice, the ratio was 0.09, which increased to 0.25 with testosterone injection.
The rate of granule-protein synthesis and total protein synthesis in normal and castrated male mice increased with 17β-estradiol injection significantly. However, the ratio did not change with 17β-estradiol as opposed to testosterone. This indicates that testosterone stimu-lates granule-protein synthesis to a much greater extent than total protein synthesis with castrated male and normal female mice, whereas 17β-estradiol stimulated both syntheses in the same manner with normal male, castrated male and normal female mice. Probably, testosterone has a stimulative effect specific for granule-protein synthesis. It seems likely that the ratio reflected the action of male hormones more precisely.View full abstractDownload PDF (579K) -
A Possible Endogenous Probe of Conformational Changes in the Mitochondrial MembraneOsamu HATASE, Ken TSUTSUI, Takuzo ODA1977 Volume 82 Issue 2 Pages 359-363
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe protein-bound sulfhydryl (SH) groups of the mitochondrial membrane were determined with Ellman's reagent in energized and non-energized configurational states of mitochondria and submitochondrial particles. When beef heart mitochondria were energized by respiration, there was a decrease in titratable protein-bound SH groups which varied according to substrate: NADH-linked substrates induced a decrease of about 10 nmol per mg of protein, succinate about 7, and ascorbate-tetramethyl-p -phenylene-diamine about 3. Similar changes occurred in phosphorylating submitochondrial particles. A decrease in SH titer was also observed in non-energized conditions, induced by hypotonic treatment and by some reagents inhibiting electron transport and oxidative phosphorylation and inducing orthodox configuration. These changes in protein-bound SH groups might be useful in analyzing the conformational states of mitochondrial membranes.View full abstractDownload PDF (366K) -
Fumio SAKIYAMA1977 Volume 82 Issue 2 Pages 365-375
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSConditions for the selective cleavage of the kynurenyl bond in oxidized hen egg-white lysozyme[EC 3. 2. 1. 17] in aqueous hydrazine solution were established at around pH 5.8. The cleavage reaction was best performed at room temperature or 30°C by direct reaction of oxidized lysozyme with 2M hydrazine containing 8 M urea or 5M guanidine hydrochloride at the optimum pH for 4 days or by reaction in the same medium for 24h after anhydrous hydrazine treatment.
The reduced and S-carboxymethylated (Rcm) derivative of an inactive oxidized lysozyme containing one N'-formylkynurenine residue (1-NFK-lysozyme) was subjected to reaction with hydrazine under the established conditions, and was split into two polypeptides. These polypeptides were isolated by gel-filtration on Sephadex G-50. Amino acid compositions and terminal residues of these polypeptides were analyzed, and the formation of 4-acylamino-6-(2'-aminophenyl)-2, 3, 4, 5-tetrahydropyridaz-3-one upon hydrazinolysis of the N'-formylkynurenyl bond was tested. These analyses showed that one of the two isolated polypeptides was the N-peptide corresponding to residues I to 62 and the other was the C-peptide consisting of residues 63 to 129 in Rcm-l-NFK-lysozyme. It was deduced that N'-formylkynurenine in I-NFK-lysozyme is present at position 62 in place of tryptophan in the native protein.View full abstractDownload PDF (836K) -
The Complete Amino Acid Sequence of COOH-Terminal Portion Following the Bradykinin MoietyHisao KATO, Yong Nam HAN, Sadaaki IWANAGA1977 Volume 82 Issue 2 Pages 377-385
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe complete amino acid sequence of the COOH-terminal portion following the bradykinin moiety in bovine plasma low-molecular-weight (LMW) kininogen was determined. The COOH-terminal segment, which corresponds to the light chain of kinin-free protein derived from LMW kininogen, consisted of a total of 47 amino acid residues with a NH2-terminal serine and COOH-terminal alanine. The sequence studies of the whole segment and the fragments obtained from the digests with trypsin [EC 3. 4. 21. 4], thermolysin [EC 3. 4. 24. 4], and Staphylococcal protease showed the following sequence:
H-1 Ser-Val-Gln-Val-Met-Lys-Thr-Glu-Gly-10 Ser-Thr-Thr-Thr-His-Val-Lys-Ser-Cys-Glu-20 Tyr-Lys-Gly-Arg-Pro-Gln-Glu-Ala-Gly-Ala-30 Glu-Pro-Ala-Pro-Gln-Gly-Glu-Val-Ser-Leu-40 Pro-Ala-Glu-Ser-Pro-Gl n-Leu-Ala-OH
The first five-NH2-terminal sequence of the segment was identical to the COOH-terminalsequence of the cyanogen bromide-fragment (Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-Ser-Val-Ghr-Val-Met) derived from LMW kininogen. This indicates the peptide linkage between the kinin moiety and the COOH-terminal segment in the parent molecule. Furthermore, the first twelve-NH2-terminal sequence was the same as the amino acid sequence of fragment 1•2, which is in the region following the kinin moiety in high-molecular-weight (HMW) kininogen. However, no such sequence homology was found in the subsequent residues up to the COOH-terminal end of LMW kininogen. These results provide further evidence that the COOH-terminal structures of LMW and HMW kininogens are different.View full abstractDownload PDF (566K) -
Grace Y. HAN, Peter F. HAN, Scott OWEN, Joe JOHNSON, Jr.1977 Volume 82 Issue 2 Pages 387-394
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe extreme sensitivity of chicken muscle fructose 1, 6-bisphosphatase to inhibition by 5'-AMPhas been utilized to develop a new method for the assay of cAMP phosphodiesterase activity. In this method, the substrate (cAMP) is first incubated with phosphodiesterase and the amount of 5'-AMP formed is then determined by measuring the degree of inhibition of fructose 1, 6-bis-
phosphatase activity. The present method conveniently employs the spectrophotometric technique and is sensitive enough to detect the conversion of 50 pmol of cAMP to 5'-AMP in 1 ml of reaction mixture. This method is considered particularly valuable for those laboratories that are not equipped with facilities for measuring radioactivity.View full abstractDownload PDF (596K) -
Isamu SHIIO, Hachiro OZAKI, Kyoko UJIGAWA1977 Volume 82 Issue 2 Pages 395-405
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSCitrate synthase [EC 4. 1. 3. 7] was purified about 50-fold from the sonicate of Brevibacterium flavum, a glutamate-producing bacterium. The purified enzyme was irreversibly inactivated in low ionic strength buffer but was stabilized by the addition of inorganic salts or of oxalacetate, a substrate of the enzyme. The enzyme had a molecular weight of 92, 000 and an optimum pH of 8.0. The enzyme was activated about 10-fold by 50mM monovalent cations (Li+, Na+, K+, NH4+, Tris+) as well as by 5mM divalent cations (Mg2+, Cat2+).
Kinetic analysis indicated that the enzymatic reaction proceeded by a rapid equilibrium random Bi Bi mechanism with two independent substrate-binding sites for oxalacetate and acetyl-CoA. The Michaelis constants for oxalacetate and acetyl-CoA were 3.9μM and 54μM, respectively, and the dissociation constants for the reaction products, citrate and CoA, were 4.5mM and 0.21mM, respectively.
The enzyme was specifically inhibited by ATP or by cis-aconitate and isocitrate but not by 2-oxoglutarate or NADH. No synergistic interaction between ATP and cis-aconitate was oberved. The extent of inhibition by 5mM cis-aconitate was about 90 per cent at pH 7.0 and 50 per cent at pH 8.0; it was partially competitive with respect to oxalacetate and non-competitive with respect to acetyl-CoA. The inhibitor constant for cis-aconitate was 0.31mM at pH 7.0 and 0.79mM at pH 8.0. Similar cis-aconitate inhibition was observed with citrate synthases of Escherichia coli, Bacillus subtilis, and pig heart.
ATP inhibited the enzyme about 50 per cent at a concentration of 5mM and showed cooperativity at lower concentrations. The inhibition was partially competitive with respect to acetyl-CoA and non-competitive with respect to oxalacetate. The inhibitor constant for ATP was 1.9mM. Based on these results, a sequential feedback control mechanism including cis-aconitate inhibition of citrate synthase was proposed for glutamate biosynthesis in B. flavum.View full abstractDownload PDF (730K) -
Taro HAYAKAWA, Yasushi MYOKEI, Haruhiko YAGI, Donald M. JERINA1977 Volume 82 Issue 2 Pages 407-415
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSGlutathione-S-epoxide transferase has been purified from guinea pig liver in a homogeneous form having a sedimentation coefficient, s20, w of 3.8S, and a molecular weight of 46, 000, and dissociable into subunits with a molecular weight of 25, 000. The activities with four substrates, styrene oxide, naphthalene oxide, iodomethane, and p-nitrobenzyl chloride, were all copurified during purification of the enzyme from 100, 000×g supernatant of guinea pig liver. However, kinetic data suggest different mechanisms for the glutathione conjugating reaction with iodomethane and p-nitrobenzyl chloride. The view that the epoxide transferase does not distinguish between simple epoxides and arene oxides has been confirmed with the guinea pig liver enzyme.View full abstractDownload PDF (1940K) -
Haruko FUJIMORI, Masatake OHNISHI, Mitsuaki SAKODA, Ryuichi MATSUNO, K ...1977 Volume 82 Issue 2 Pages 417-427
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe enzymatic reaction catalyzed by saccharifying α-amylase from Bacillus subtilis was studiedkinetically by following the reaction using the polarimetric method and quantitative paperchromatography. The time course of the reaction with purified maltose as a substrate showed a distinct lag phase in the initial stage of the reaction, followed by a linear steady-state phase. The steady-state rate showed a strong sigmoidal dependency on the concentration of substrate m altose, as was previously reported for the initial rate by Shibaoka et al. (Shihaoka, T., Inatani, T., Hiromi, K., and Watanabe, T., J. Biochem., 77, 965-968 (1975)). The initial lag phase can be eliminated by the addition of a small amount of maltotriose.
Quantitative paper chromatographic determination of the substrate and products during the course of the reaction showed no detectable change in the lag phase, after which maltotriose and altotetraose appeared in the steady-state and finally disappeared. These results clearly indicate that the reaction involves glycosyl transfer and/or condensation in addition to hydrol ysis and that maltotriose and maltotetraose formed strongly accelerate the reaction. To interpret these phenomena, a reaction scheme involving hydrolysis, glycosyl transfer and condensation is presented. Based on this scheme, a computer simulation of the time course of the reaction was attempted.
Computer simulation based on this scheme and the subsite structure of the enzyme reproduced the lag phase and the strong sigmoidal concentration dependence of the rate, confirming the validity of the reaction mechanism proposed.
The major factors responsible for the lag and the sigmoidicity are as follows: (i) The glucosidic linkage in maltose is difficult to cleave either for hydrolysis or glycosyl transfer. (ii) The rate constant of glycosyl transfer is much larger than that of hydrolysis. (iii) Maltotriose and maltotetraose are much better substrates than maltose in hydrolysis and/or in glycosyl transfer. (iv) Once these maltooligosaccharides are formed from maltose, even in a small amount, by transglycosylation or condensation, a reaction cycle is formed by which glycosyl transfer and hydrolysis are greatly accelerated. In this cycle maltose is consumed as an acceptor and glucose is formed as a product split off either in the hydrolysis or in the glycosyl transfer. This mechanism, together with the subsite structure, also explains why this enzyme yields glucose in much higher amount than other α-amylases.View full abstractDownload PDF (1300K) -
III. Ionic Strength Effects and Kinetics in the Action of Chondroitinase ACKeiichiro HIYAMA, Shigetaka OKADA1977 Volume 82 Issue 2 Pages 429-436
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSIn order to compare the action of Arthrobacter chondroitinase AC [EC 4. 2. 2. 5] (A-Chase) with that of Flavobacterium chondroitinase AC (F-Chase), steady-state kinetic studies were carried out at pH 6.0. Preliminary experiments showed the action of A-Chase to be accelerated by the addition of a salt to the reaction mixture. The degree of activation by the salt additive was maximum at the ionic strength μ=0.05. The action of A-Chase was suppressed by the salt at ionic strength higher than 0.05. Several bivalent metal cations such as Ca2+, Co2+, Mg2+, and Ba2+ were more effective for activation than univalent metal cations such as Na+, K+, etc. Further, A-Chase was inactivated by Pb2+, Cu2+, Fe2+, and Sn2+. Kinetic studies on the effect of ionic strength showed that an ionic additive causes both activation of the substrate and inhibition of A-Chase. It was also shown that DS is a competitive inhibitor of A-Chase; the dissociation constant of the enzyme-inhibitor complex, K1, is 6.1×10-5M as moles of repeating disaccharide unit. Heparin is not an inhibitor of the enzyme.
Kinetic parameters of the action of A-Chase and F-Chase were estimated for ChS-A, ChS-C, Ch, and HA. Van't Hoff plots of the apparent Km. values and Arrhenius plots of the apparent Ko values for both enzymes with these substrates gave linear relationships with the exception of the plots of Ko for F-Chase. The thermodynamic and activation parameters under 30°C were calculated assuming a Michaelis-Menten type mechanism with the enzyme and the substrate in rapid equilibrium. The ΔH values obtained for various substrates were negative for A-Chase and positive for F-Chase.
The results obtained in this study suggest that the driving force of enzyme-substrate com-plex formation can be attributed to ionic interaction for A-Chase and to hydrophobic bonding for F-Chase.View full abstractDownload PDF (524K) -
Yoshimi OTA, Ken-ichi ARAI, Yoshito KAZIRO1977 Volume 82 Issue 2 Pages 437-447
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe polypeptide chain release factor 1 (RF-1) has been purified from an extreme thermophile, Thermus thermophilus HB8. The purification procedure included steps of aqueous two-phase partition, ammonium sulfate fractionation, and column chromatographies on DEAE-Sephadex, Sephadex G-150, and CM-Sephadex. The preparation was more than 90% pure as judged by polyacrylamide gel electrophoresis. The specific activity was about 3.3 pmol of formyl-[3H] methionine released in 1 min at 25°C per μg of protein under the standard assay conditions using 4 pmol of the initiation complex and 1 nmol of UpApG. The molecular weight as determined by gel filtration on Sephadex G-150 and by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, was 58, 000 and 45, 000, respectively. As expected, the factor was extremely heat-stable, 50% of its activity remaining after incubation for 5min at 84°C. Several properties of the reaction catalyzed by RF-1 are also described.View full abstractDownload PDF (828K) -
Fumio MIAKE, Takao TORIKATA, Katsumi KOGA, Katsuya HAYASHI1977 Volume 82 Issue 2 Pages 449-454
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSNADP+-specific isocitrate dehydrogenase was found in several tissues of the pupa of the silkworm, Bombyx mori. This enzyme was highly purified from the whole bodies of pupae. This is the first isolation of the enzyme from insect materials. The purified enzyme gave a single protein band on polyacrylamide gel electrophoresis. The reaction catalyzed by the purified enzyme was readily reversible. The pH optimum for the forward reaction (reduction of NADP+) was 7.8, and that for the reverse reaction (oxidation of NADPH) was 6.6. The enzyme had a molecular weight of 86, 000 and was found to be composed of two identical subunits, which have a molecular weight of 44, 000. The activity of the enzyme in the forward reaction was slightly inhibited by citrate, oxaloacetate, α -ketoglutarate, and others. Citrate stabilized the activity over a wide pH region.View full abstractDownload PDF (452K) -
Masamichi TAKAGI1977 Volume 82 Issue 2 Pages 455-460
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSTo assess the effect of alteration of membrane structure on the enzymic activities related to phospholipid synthesis in microsomal membrane, the effects of several organic solvents have been studied in an in vitro system, in which the cytoplasmic extract prepared from rat liver incorporated [14C] choline or [14C] CDP-choline into phosphatidylcholine (lecithin). The optimum conditions for the incorporation were determined.
Among several organic solvents examined, n-alkanes such as n-hexane, n-octane, and n-tetradecane stimulated the incorporation.
It was shown that n-alkanes stimulated one of three enzymic steps of lecithin biosynthesis from choline; that is, the formation of CDP-choline catalyzed by CTP: cholinephosphate cytidyltransferase [EC 2. 7. 7. 15], an enzyme on the microsomal membrane. It was further shown that the same enzyme was also stimulated by preincubation of microsomes in the absence of substrate.
It is suggested that alteration of the lipid environment of the microsomal membrane induced by n-alkanes caused activation of this enzymic step.View full abstractDownload PDF (423K) -
Akemi SUZUKI, Mayumi SATO, Shizuo HANDA, Yasutoshi MUTO, Tamio YAMAKAW ...1977 Volume 82 Issue 2 Pages 461-467
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSChanges of seminolipid content in vitamin A-deficient rat testes were quantitatively determined by improved and facile high performance liquid chromatography using a reversed phase column under an ion pair chromatographical condition. The seminolipid content of the testes of rats fed a vitamin A-deficient diet for 46 days decreased to 13% of that of the control rats fed a vitamin A-deficient diet for 20 days and then supplemented with 140μg/rat/day of vitamin A palmitate for 26 days. Total lipid, phospholipid and DNA of vitamin A-deficient rats were slightly reduced. Histological examinations showed that seminiferous tubular cells degenerated to aspermatogenesis with a vitamin A-deficient diet.
The remarkable reduction of seminolipid content, compared to the slight decrease of DNA content, is considered to be the result of damage to the seminiferous tubular elements, especially at the differentiated step of germinal cells from spermatogonia to the next. These results further support the theory that seminolipid biosynthesis is characterized by strict dependence on the differentiation of seminiferous tubular cells or spermatogenesis.View full abstractDownload PDF (969K) -
Koki UCHIDA, Umeji MURAKAMI, Toshiaki HIRATSUKA1977 Volume 82 Issue 2 Pages 469-476
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSUsing phenylmethanesulfonyl fluoride (PMSF) in all the steps of the preparation procedure to inhibit protease activity, purified cardiac myosin was isolated from rat hearts. The purified myosin obtained was free from nucleic acid, actin, tropomyosin, C-protein, and proteolytic products of myosin. Without PMSF, the myosin preparation containing proteolytic products was obtained.
The properties of the ATPase of rat cardiac myosin [EC 3. 6. 1. 3] were studied. The Ca2+-, K+-, and Mg2+-ATPase activities of the purified myosin at high ionic strength and pH 7.0 were 1.15, 0.2, and 0.05μmol P1mg-1min-1, respectively. Those of the myosin prepared without PMSF were lower than the above values and fluctuated from preparation to preparation. The Ca2+-ATPase activity of rat cardiac myosin was not activated by PCMB, although the content of SH groups was 6.6-7 mol per 105g myosin. This anomalous property and the content of SH groups were also found in the myosin preparation containing proteolytic products.
Analysis of subunits in the purified myosin was carried out on 3.5% acrylamide gels with 0.1% SDS. Of the total protein present in myosin, 10.2% was in the light chains; LCI, 6.2% and LC2, 4.0%. Urea gel electrophoresis of rat cardiac myosin showed a band correspondingto LC1 and two bands coresponding to LC2.
Without the protease inhibitor, rat cardiac myosin underwent proteolytic cleavage duringits preparation, producing two fragments with molecular weights of 117, 000 and 160, 000. It is thought that one of them, the 117, 000 fragment, is a constituent of subfragment 1 produced from cardiac myosin by inherent protease activity in rat heart cells.View full abstractDownload PDF (2380K) -
Taishin TAKUMA, Masayoshi KUMEGAWA1977 Volume 82 Issue 2 Pages 477-481
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAn increase of thymidine kinase [EC 2. 7. 1. 21] activity and decrease of 5'-nucleotidase [EC 3. 1. 3. 5] activity for dTMP were found during hormonal regeneration of the seminal vesicles by daily or single administration of testosterone propionate into mice castrated 2 weeks previously. Actinomycin D injected on day 0 of testosterone treatment completely inhibited both the increase of thymidine kinase and the decrease of 5'-nucleotidase. When injected on day 2. actinomycin D decreased thymidine kinase activity below the control level and 5'-nucleotidase activity was not restored to the normal level. The activity of 5'-nucleotidase in a mixed sample, in which seminal vesicles of castrated mice and those of testosteronetreated mice were homogenized together, was intermediate between the activities determined separately. This indicates the absence of any inhibitor of 5'-nucleotidase in the regenerating vesicles. Changes in total activity of 5'-nucleotidase and total protein content in extracts during various treatments showed that the decrease in specific activity of 5'-nucleotidase in the first 2 days of testosterone treatment was not due to inhibition of enzyme activity but to dilution of the enzyme with other proteins which increased in content more rapidly than 5'-nucleotidase.View full abstractDownload PDF (379K) -
Kazumori MASAMOTO, Mitsuo NISHIMURA1977 Volume 82 Issue 2 Pages 483-487
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSCarbonylcyanide m-chlorophenylhydrazone (CCCP) or nigericin induced translocation of H+ in the dark across the cell membrane of blue-green algae Plectonema boryanum and Anacystis nidulans. The direction of the H+ flux depended on the pH of the suspending medium. At acidic pH, an influx of H+ and at alkaline pH an efflux of H+ were observed. It is suggested that the influx takes place at pH's higher than the “internal” pH and the efflux at pH's lower than that. The internal pH was estimated to be 7.4±0.2 for Plectonema boryanum and 7.5±0.1 for Anacystis nidulans. Similar H+ changes due to CCCP were observed under illumination, where the light-induced efflux of H+ was limited by the counter-flux of cations. The internal pH of cells in the light, estimated from the pH-dependent reversion in the rate of the H+ change, was about 8.5.View full abstractDownload PDF (380K) -
Difference between Isotonically and Hypotonically Treated Preparations and Effects of Divalent CationsKazumori MASAMOTO, Mitsuo NISHIMURA1977 Volume 82 Issue 2 Pages 489-493
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSLysozyme-treated cells of a blue-green alga, Plectonema boryanum, had an internal pH of 7.3±0.2 under isotonic and hypotonic conditions. This value was similar to that of untreated cells. The CCCP-induced biphasic H+ change seen in the isotonic cells was not observed in the hypotonically treated cells. The biphasic time course remained in the hypotonic preparation if CaCl2 or MgCl2 was added prior to the osmotic shock. It is suggested that the cells have two compartments of H+ concentration. The outer region may be more acidic than the inner region.
A light-induced H+ efflux was observed under isotonic conditions and an influx of H+ under hypotonic conditions. The H+ influx was not observed when lysozyme-treated cells were incubated with CaCl2 or MgCl2 prior to the hypotonic treatment. Two types of effects of divalent cations, one on the rigidity of the outer membrane and another on the permeability characteristics of the inner photosynthetic membrane, are indicated. Rearrangement of the photosynthetic membranes and an apparent inversion of the H+ pump by hypotonic shock are also suggested.View full abstractDownload PDF (372K) -
Akihiro YOSHIDA, Tetsu HOZUMI, Katsuhisa TAWADA1977 Volume 82 Issue 2 Pages 495-498
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe isometric tension of single fibers isolated from glycerinated rabbit psoas muscle was measured at various temperatures using Mg-ITP as a substrate. The tension developed in Mg-ITP decreased linearly as the temperature was reduced from 24°C to 4°C.
Myosin formed the myosin*-product complex predominantly via ATP hydrolysis at the burst site during Mg-ATP hydrolysis, irrespective of temperature, and the tension developed in Mg-ATP decreased linearly as the temperature decreased (Yoshida and Tawada (1976) J. Biochem. 80, 861). During Mg-ITP hydrolysis, myosin forms the myosin*-product complex predominantly at the burst site above 20°C, while myosin forms the myosin*-substrate complex below 8°C (Hozumi (1976) Eur. J. Biochem. 63, 241). However, the temperature dependence of tension development in Mg-ITP is linear, as with Mg-ATP, as mentioned above. This temperature dependence is not compatible with some muscle models which assume the formation of the myosin*-product complex by cross-bridges prior to combination with actin during contraction.View full abstractDownload PDF (225K) -
Hiroshi HATANAKA, Fujio EGAMI1977 Volume 82 Issue 2 Pages 499-502
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAmino acids produced from formaldehyde and hydroxylamine in modified sea mediums with different concentrations of molybdate were analyzed. The modified sea mediums contained lower concentration of sodium chloride and higher concentrations of transition metal ions (Zn2+, Fe3+, Cu2+, Co2+, Mn2+ each 10-4M, and MoO42- 10-6, 10-4, or 10-2M) than sea water.
The concentration of molybdate had apparently no remarkable effect on the total yields of primary amino groups, but a remarkable effect on the nature of amino acids produced. The formation of alanine, aspartic acid, β-alanine and, in particular, proline was increased, and that of glycine and serine was decreased with the enrichment of molybdate.
The results suggest the possibility of a natural selection of prebiotic organic molecules based on the nature of environmental catalysers in the course of chemical evolution.View full abstractDownload PDF (272K) -
Variable and Conserved Regions in the SequencesHiroaki HAYASHI, Koichi IWAI, Jerry D. JOHNSON, James BONNER1977 Volume 82 Issue 2 Pages 503-510
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSPea histone II group, a mixture of H2A and H2B obtained by chromatography on an ionexchange resin, was further fractionated by carboxymethylcellulose chromatography and purified by Bio-Gel P-60 chromatography. Their chromatographic behaviors and gel electrophoretic mobilities of single bands differed significantly from those of calf H2A and H2B. Their amino acid compositions were similar to those of the calf histones as a whole, but differed in detail in certain respects. The partial sequence of pea H2B was deduced from the amino acid compositions of BrCN cleavage fragments and tryptic peptides in comparison with the known sequence of calf H2B. It is different in the amino-terminal basic region from the calf H2B, with a blocked amino terminal and a larger number of residues. In contrast, the middle and carboxy-terminal hydrophobic regions are relatively similar, with at least 19-21 different residues and microheterogeneity at two positions of the pea sequence. The sequence of H2A may vary in much the same way as that of H2B, as suggested by the similar extent of differences in their amino acid compositions. It is thus assumed that the amino-terminal regions, at least, of H2A and H2B histones are variable in evolution provided that they remain basic enough to bind DNA, whereas the middle and carboxy-terminal hydrophobic regions of H2A, H2B, H3, and H4 should be conserved to ensure precise histone core formation inside the repeated units of chromatin.View full abstractDownload PDF (483K) -
Haruki YAMAGUCHI, Kenji OKAMOTO1977 Volume 82 Issue 2 Pages 511-518
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe reducing-end sugar residues of pachyman, amylose, dextran, and cold water-insoluble laminaran were isolated from the polymers as penta-O-methylhexitols or penta-O-ethylhexitols by treatment with a strongly basic anion-exchange resin (OH- form) after reduction, alkylation, and acid hydrolysis. The penta-O-alkylhexitols obtained were analyzed by GLC and combined GLC-mass spectrometry as acetates and found to be in accord with expectations based on the known structures of the polysaccharides. 3- and 4-mono-O-acetyl-penta-O-alkylhexitols, which have the same substitution pattern, were easily distinguishable on the mass spectrograms, using sodium borodeuteride for reduction.View full abstractDownload PDF (524K) -
Hiroshi YANAGAWA, Yoko OGAWA, Fujio EGAMI1977 Volume 82 Issue 2 Pages 519-528
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSTwo novel deoxyribonucleases, termed DNases A and A', have been purified from the hepatopancreas of Achatina fulica (agate snail). DNases A and A' were obtained in 3.6% and 7.7% yields by acetate buffer extraction and successive chromatography on hydroxyapatite, phosphocellulose and poly (A)-Sepharose. The two DNases are highly active toward poly (dA) and at a salt concentration of 0.15M and pH 5.0 exhibit 10-fold higher hydrolyzing activities toward poly (dA) than toward calf thymus denatured DNA. The enzymes also act considerably on poly (dT) at pH 4.0, but do not appreciably digest other synthetic homopolymers such as poly (dC), poly (dG) and poly (A). The limit products obtained by exhaustive digestion of poly (dA) with DNases A and A' are 98% and 99% acid-soluble and consist of oligonucleotides with an average chain length of about 5 nucleotides containing barely detectable dimers and trimers, respectively. These fragments have terminal 5'-hydroxyl and 3'-phosphate groups. The mode of action appears to be endonucleolytic. Both enzymes have the same pH optimum of 5.0 for poly(dA)hydrolyzing activity. Ionic strength is critical for the maximum activity.View full abstractDownload PDF (701K) -
Toshiharu MURAOKA, Hiromichi OKUDA1977 Volume 82 Issue 2 Pages 529-533
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSA novel method for the estimation of fibrinolytic activity is proposed. In this method, a fibrin clot suspension is used as a substrate (fibrin is known to be a physiological substrate of plasmin). The fibrin clot suspension was prepared by homogenization of human fibrin clots. With this method, we found that free fatty acids inhibited the plasmin activity, and long-chain, unsaturated free fatty acids had a particularly strong inhibitory action on plasmin. As regards the mechanism of the inhibitory action, free fatty acids may not inhibit complex formation between plasmin and fibrin, but may make it impossible for plasmin to act on fibrin due to deformation of the surface of the fibrin clot.View full abstractDownload PDF (360K) -
Properties of the Bound Pyridoxal 5'-PhosphateHaruo MISONO, Kenji SODA1977 Volume 82 Issue 2 Pages 535-543
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSL-Lysine-α-ketoglutarate ε-aminotransferase of Flavobacterium lutescens (Achromobacter liquidum) IFO 3084 shows positive circular dichroic bands at 340 and 415nm where absorption maxima are observed, and fluorescence maxima at 380 and 490nm on excitation at 340 and 415nm, respectively. The pyridoxal 5'-phosphate absorbing at 415nm is bound through an aldimine linkage to an ε-amino group of the lysine residue of the protein. Upon aging, the 415nm pyridoxal 5'-phosphate changes to a less active form (λmax, 325nm), which is distinguishable from the 340nm pyridoxal 5'-phosphate. This 325nm bound pyridoxal 5'-phosphate is reduced with sodium borohydride and shows no circular dichroism. When the semiapoenzyme is aged under the same conditions, no spectral change is observed. These findings suggest that the pyridoxal 5'-phosphate bound through an aldimine linkage may be converted into a carbinol amine or some other related form by aging.View full abstractDownload PDF (618K) -
Yoshinori ENDO, Katsuko YAMASHITA, Yong Nam HAN, Sadaaki IWANAGA, Akir ...1977 Volume 82 Issue 2 Pages 545-550
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSFragment 1, released from bovine plasma high-molecular-weight kininogen by the action of plasma kallikrein, is a glycopeptide with approximately 3 mol of each of N-acetylgalactosamine, galactose and sialic acid in one molecule. All these sugars were in the T-1 fragment obtained by tryptic digestion of fragment 1. Sialic acid can be completely released from the T-1 fragment by sialidase digestion. When this sialic acid-free T-l fragment was incubated with purified diplococcal endo-α-N-acetylgalactosaminidase, all remaining sugars were released as a disaccharide. By Smith degradation and β-galactosidase digestion, the structure of this disaccharide was found to be Gal β1→3Ga1NAc.
Methylation analysis of the trisaccharide released from fragment T-1 by alkaline borohydride treatment indicated that all the galactose was obtained as the 2, 4, 6-tri-O-methyl derivative. Based on this evidence, the complete structure of the carbohydrate moieties of fragment 1 was proposed as Sialylα2→3Galβ1→3GalNAc. In addition, small amounts of a tetrasaccharide, Sialylα2→3Galβ1→3 (Sialylα2→6) GalNAc also occurred as a carbohydrate chain of fragment 1.View full abstractDownload PDF (438K) -
Changes of Ornithine and Acetylglutamate Concentrations in the Livers of Rats Subjected to Dietary TransitionsTakeyori SAHEKI, Tsunehiko KATSUNUMA, Mariko SASE1977 Volume 82 Issue 2 Pages 551-558
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSRegulation of urea synthesis in vivo was studied in rats subjected to acute dietary transitions from 70% to 5% casein diet or vice versa.
Significant increase/decrease of urea excretion and urea in the liver preceded increase/decrease of argininosuccinate synthetase [EC 6. 3. 4. 5] activity during the dietary transitions.
Immunochemical determination of argininosuccinate synthetase showed that the increase/ decrease of the enzyme activity is directly related to the actual increase/decrease of the amount of enzyme protein.
The ratios of the initial velocity of urea synthesis from ammonium chloride to the enzymatic activity of argininosuccinate synthetase in the perfused livers were determined. Shortly after the switch of casein diet from 70% to 5%, the ratios decreased to much lower values than those of rats fed on the 5% casein diet for 7 days, while, during the acute transition from 5% to 70% casein diet, the ratios increased above the values of rats fed on the 70% casein diet for 7 days.
The concentrations of ornithine and acetylglutamate capable of stimulating urea synthesis changed significantly shortly after the dietary switch. In particular, the concentration of ornithine overshot the steady-state level of rats fed on 70% casein after the dietary switch from 5% to 70% casein. Ornithine concentration declined rapidly after the dietary switch from 70% to 5%, falling below the steady-state level of rats fed on the 5% casein diet. Thus, in both cases, new steady-state levels of ornithine concentration were established. The findings that fluctuations of ornithine and acetylglutamate concentration were greater and occurred prior to the activity changes of argininosuccinate synthetase strongly suggested a possible regulatory function of these amino acids in urea synthesis.View full abstractDownload PDF (618K) -
A Kinase Activity for D-Glucosamine without Inhibition by Glucose in Rat Liver and Its Possible Identity with GlucokinaseMichihiko OGUCHI, Machiko SATO, Yoko MIYATAKE, Nobu AKAMATSU1977 Volume 82 Issue 2 Pages 559-567
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSPhosphorylation of D-glucosamine with rat liver extracts and the effects of fasting and partial hepatectomy on its activity were studied. While the phosphorylation with brain extract is greatly inhibited by glucose and glucose-6-phosphate, the activity of liver extract has a low sensitivity to these compounds. This kinase activity as well as glucokinase [EC 2. 7. 1. 2] decreased when animals were fasted for 48 to 72h, whereas no appreciable changes were found in low Km hexokinase [EC 2. 7. 1. 1] activity. Studies on DEAE-cellulose column chromatography showed that glucosamine was phosphorylated not only by low Km hexokinase but also by the kinase corresponding to glucokinase. The latter enzyme peak on chromatography was reduced by fasting for 72h to half that of normal animals. The activity of phosphorylat ion of glucosamine associated with glucokinase was also reduced by the same proportion. Both activities were restored almost to those of normal liver by refeeding. Partial hepatectomy had similar effects to fasting. The activity of phosphorylation of glucosamine as well as glucokinase activity fell to 45% of the unoperated controls 48h after the operation, whereas low Km hexokinase activity remained unchanged.
These results substantiate the finding that glucosamine is phosphorylated to an appreciable extent by hepatic glucokinase, an enzyme previously believed not to phosphorylate this hexosamine. The apparent Km value of 8mM for glucosamine was lower than that for glucose (10-20mM). While phosphorylation of glucosamine by low Km hexokinase was greatly inhibited by glucose, the activity associated with glucokinase was not sensitive to glucose up to 40mM.View full abstractDownload PDF (732K) -
Hitoshi AOSHIMA1977 Volume 82 Issue 2 Pages 569-573
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSSeparation of 13-hydroperoxylinoleic acid or 13-hydroperoxylinolenic acid from linoleic acid or linolenic acid, respectively, was carried out easily and quickly by high performance liquid chromatography on porous polymer gel (TSK-Gel LS-140) using n-hexane/ethanol as an eluent. An eluent containing a large amount of n-hexane (96%) made possible the separation of 9- and 13-hydroperoxylinoleic acids. These methods were applicable for analyses of the products obtained by the incubation of soybean lipoxygenase-1 [linoleate: oxygen oxidoreductase, EC 1. 13. 11. 12] with linoleic acid or 13-hydroperoxylinoleic acid.View full abstractDownload PDF (281K) -
Toshiaki HIRATSUKA, Koki UCHIDA, Koichi YAGI1977 Volume 82 Issue 2 Pages 575-583
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. An initial burst of P1 liberation (1.2 mol per mol of heavy meromyosin) was observed in the hydrolysis of 2' (or 3')-O-(2, 4, 6-trinitrophenyl)adenosine 5'-triphosphate (TNP-ATP) by heavy meromyosin ATPase [EC 3. 6. 1. 3]. When heavy meromyosin was preincubated with excess TNP-ATP, the size of the initial burst of P1 liberation from the added ATP increased with ATP concentration and reached a constant level (1.28 mol) at a 1.9 molar excess of ATP to the enzyme.
2. On preincubation of heavy meromyosin with excess TNP-ATP, the steady-state ATPase activity was reduced to below 20% of the control at low ATP concentrations (below 0.5mM) without change in the size of the initial burst of P1 liberation. However, it recovered with increase of ATP concentration and reached the same level as the control at higher ATP concentrations (above 1mM). The difference spectrum at 518 run, which is induced by TNP-ADP interacting with a tryptophanyl residue of heavy meromyosin, decreased with increase of concentration of ATP added and almost disappeared at 0.9mM ATP, where the steady-state ATPase activity had recovered to 90 % of the control.
3. On the reaction of heavy meromyosin with TNP-ATP, a complex of heavy meromyosin with 2 mol of TNP-ADP was isolated by dialysis. One mole of TNP-ADP was readily released on incubation with 0.2-0.4mM ATP or by ammonium sulfate precipitation before dialysis. Higher concentrations of ATP (above 1mM) were necessary to release the other I mol of TNP-ADP.
4. The results of the present study show differences between the two bindings of TNP-ADP to heavy meromyosin, one to the active site for the initial burst of P1 liberation and the other to that for the steady-state ATPase. The former is readily disrupted by adding low concentrations of ATP or by precipitation with ammonium sulfate, and the latter, involving interaction with a tryptophanyl residue, is stable even in the presence of sodium dodecyl sulfate.View full abstractDownload PDF (579K) -
Seiki KURAMITSU, Kiyoshi IKEDA, Kozo HAMAGUCHI1977 Volume 82 Issue 2 Pages 585-597
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe pH difference spectra of intact hen lysozyme [EC 3. 2. 1. 17], Asp 52-esterified lysozyme, and Glu 35-Trp 108-esterified lysozyme were measured at various ionic strengths and temperatures. The pH dependence of the extinction difference at 301.5nm was well interpreted in terms of the ionization of the catalytic groups, Asp 52 and Glu 35, which interact electrostatically. The macroscopic pK values of Asp 52 and Glu 35 were determined to be 3.4 and 6.1, respectively, at 0.1 ionic strength and 25° C.
Regarding the ionization of the catalytic groups as that of a dibasic acid and assuming that the microscopic ionization constant of Glu 35 of intact lysozyme in which Asp 52 has been protonated (pk1, 35) is equal to the ionization constant of Glu 35 in Asp 52-esterified lysozyme, we estimated the four microconstants for the ionization of Asp 52 and Glu 35 in intact lysozyme. The microconstants for the first ionization of Asp 52 (pk1, 52) and Glu 35 (pk1, 35) were more sensitive to ionic strength of the medium than those for the second ionization of Asp 52 (pk2, 52) and Glu 35 (pk2, 35). The increase in pk1, 35 and pk1, 35 with an increase in ionic strength was interpreted in terms of the electrostatic interaction between the positively charged lysozyme molecule and a proton which is going to bind to Asp 52 or Glu 35, and the smaller increase in pk2, 52 and pk2, 35 in terms of an additional electrostatic interaction between negatively charged catalytic carboxylate ions. The pk values for the lysozyme molecule with zero net charge at zero ionic strength which were obtained by correcting for these electrostatic interactions were 4.7-5.1 for Asp 52 and 6.1-6.5 for Glu 35.
The absolute values of ΔH° of the ionization for Asp 52 and Glu 35 were less than 0.5 kcal•mol-1. The ΔS° values were -16 and -20 cal deg-1•mol-1 for the first and second ionizations of Asp 52, respectively, and -24 and -28 cal deg-1•mol-1 for the first and second ionizations of Glu 35, respectively, at 0.1 ionic strength and 25°C.View full abstractDownload PDF (894K) -
Akira KAMEI, Kiyoshi NAKAZAWA, Noriko TAKEUCHI, Shuzo AKASHI, Koshiro ...1977 Volume 82 Issue 2 Pages 599-602
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe presence of considerable amounts of pyruvic acid was demonstrated in the extracellular polysaccharide produced by Escherichia coli 36M isolated from a coligranuloma of chick intestine. On the basis of methylation studies and ketal exchange reaction, it was concluded that the pyruvic acid exists in the polysaccharide as 4, 6-O-(1-carboxy ethylidene)D-glucopyranoside.View full abstractDownload PDF (290K) -
Tetsuo MAITA, Etsuro TANAKA, Morris GOODMAN, Genji MATSUDA1977 Volume 82 Issue 2 Pages 603-605
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSGlobin prepared from hemoglobin of adult tupai (Tupaia glis) was separated into α and β polypeptide chains by CM-cellulose column chromatography. The S-aminoethylated α polypeptide chain and S-carboxymethylated β polypeptide chain were each digested with trypsin, and the sequences of all the peptides thus obtained were established. The ordering of these tryptic peptides in the α and, β polypeptide chains was deduced from the homology of their primary structures with that of human adult hemoglobin. In this way the primary structures of the α and β polypeptide chains of tupai hemoglobin were established; 27 amino acids in the α polypeptide chain and 26 in the β polypeptide chain differ from those in human adult hemoglobin.View full abstractDownload PDF (187K) -
VIII. Evidence for Framework Protein of the Cores of Rat Liver PeroxisomesTakafumi WATANABE, Tetsuya SUGA, Hidenori HAYASHI1977 Volume 82 Issue 2 Pages 607-609
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe cores of peroxisomes were purified 670 fold from a rat liver homogenate and the protein in the preparation was examined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis.
Two bands of protein were detected on 10% polyacrylamide gel, and their molecular weights were calculated to be about 32, 000 and 27, 000.
On treatment of the core fraction with alkali, urate oxidase was solubilized and on 10% polyacrylamide gel this fraction gave a single band of protein with an estimated molecular weight of 32, 000.
These results suggests that the protein component having a molecular weight of 27, 000 is the framework protein of the core of rat liver peroxisomes.View full abstractDownload PDF (1404K) -
Midori OGATA-ARAKAWA, Takashi MURAMATSU, Akira KOBATA1977 Volume 82 Issue 2 Pages 611-614
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAn endo-β-N-acetylglucosaminidase was isolated from fig extract. The action spectra of the partially purified enzyme on various glycopeptides showed that the substrate specificity of the enzyme is similar to that of endo-β-N-acetylglucosaminidase CII isolated from Clostridium perfringens.View full abstractDownload PDF (215K) -
Yoshiko NARAHASHI, Kaoru YODA, Shinkichi HONDA1977 Volume 82 Issue 2 Pages 615-618
Published: August 25, 1977
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSA carboxypeptidase of St. griseus K-1 (CPase S) was found to possess the specificities of both mammalian pancreatic CPase A and B. Three adsorbents for affinity chromatography were prepared by coupling L-Leu, n-Leu, and D-Arg with CH-Sepharose 4B. D-Arg-CH-Sepharose and L-Leu-CH-Sepharose retained the purified CPase S but D-Leu-CH-Sepharose did not. The activities of CPase S toward CGL and BGA were eluted in the same position. CPase S migrated as a single band on polyacrylamide gel electrophoresis and the two activities were both extracted from this band on the gel.View full abstractDownload PDF (895K)
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