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1.
Hitoshi Sawada Hideyoshi Yokosawa Shin-Ichi Ishii Motonori Hoshi 《Molecular reproduction and development》1982,5(3):291-301
The presence of a protease has been demonstrated in sperm of the solitary ascidian, Halocynthia roretzi, by using t-butyloxycarbonyl-L-Val-L-Pro-L-Arg-4-methylcoumaryl-7-amide (Boc-Val-Pro-Arg-MCA) and other arginyl or lysyl MCA derivatives as substrates. Several properties of the enzyme were investigated in a crude extract. The activity had a pH optimum near 8.0 and was enhanced by the addition of CaCl2. The Km value of 87μM was determined for Boc-Val-Pro-Arg-MCA under the optimal conditions. An apparent molecular weight was estimated to be 35,000 by gel filtration. The enzyme was inhibited with diisopropyl fluorophosphate, leupeptin, antipain, p-aminobenzamidine, Val-Pro-Arg-CH2Cl, and soybean trypsin inhibitor, but scarcely inhibited with chymostatin, elastatinal, p-chloromercuribenzoic acid, tosyl-Lys-CH2Cl, and tosyl-Phe-CH2Cl. Boc-Val-Pro-Arg-MCA, the most susceptible of the substrates examined, showed the most effective inhibition against fertilization of ascidian eggs. Thus, this enzyme in ascidian sperm extract has features closely similar to mammalian acrosin [EC 3.4.21.10], and we conclude that the enzyme is involved in fertilization as one of the lysins. 相似文献
2.
Enzymatic desulfurization of dibenzothiophene by a cell-free system of Rhodococcus erythropolis D-1 总被引:1,自引:0,他引:1
Abstract Pseudomonas syringae cells were exposed to Cu2+ alone or in the precence of acetate, proline or cysteine, at concentrations that reduced free Cu2+ to 1/10 of the total copper. Ligand concentrations (designated as isoeffective) were determined experimentally using a Cu2+ -selective electrode and confirmed by computer calculations using published stability constants. Exposure of P. syringae cells to Cu2+ alone resulted in rapid and pronounced cell death, and binding of most of the copper in solution. The addition of acetate, proline or cysteine, a few minutes after Cu2+ treatment, resulted in a significant reduction in cell death, and in the amount of copper bound to the cells. For short exposures to Cu2+ , cysteine was more effective than acetate or proline, but after 60 min of treatment, similar results were observed with these ligands. The addition of ligands before Cu2+ resulted in even more reduced copper toxicity. The results showed that, at isoeffective concentrations, weak and moderate copper-ligands can effectively antagonize copper toxicity, and that this protective effect does not require previously equilibrated copper-ligand solutions and is not very dependent of the nature of the ligand. 相似文献
3.
Junichi Ishihara Nagahiro Saijo Yasutsuna Sasaki Hidehiko Nakano Akira Ozaki Hidenobu Takahashi Masanori Sakurai Kazuhiko Nakagawa Masaaki Iigo Fumihiko Kanzawa Akio Hoshi Weon Seon Hong James R. Jett Terumi Takahashi 《Cancer immunology, immunotherapy : CII》1987,24(3):185-189
Summary The antitumor effect of recombinant human tumor necrosis factor (rH-TNF) on two clones of rat fibrosarcoma with different metastatic potential to lymph nodes was examined. The colony formation of clone A, which has high metastatic potential, was completely inhibited by continuous exposure to rH-TNF at 50 U/ml. In contrast, colony formation of clone G, which has low metastatic potential, was not inhibited by high concentrations of rH-TNF (10,000 U/ml). The inhibitory effect of rH-TNF on colony formation by clone A was also observed with a 1-h exposure to rH-TNF. This effect was time and concentration dependent, as determined by the colony assay, 3H-thymidine uptake assay, and 51Cr-release assay. 3H-thymidine and 3H-uridine uptake per cell of clone A exposed to rH-TNF was not decreased. This suggests that the mechanisms of the antitumor effect of rH-TNF were not due to inhibition of DNA and RNA synthesis of tumor cells. In vivo growth and lymph node metastases of clone A inoculated i.p. to Donryu strain rats were completely suppressed by 14 consecutive i.p. injections of 105 or 106 U/kg per day of rH-TNF. On the other hand the growth of clone G was not influenced by rH-TNF administration. 相似文献
4.
Yoshikazu Izumi Pijush Kanti Nath Hiroshi Yamamoto Hideaki Yamada 《Applied microbiology and biotechnology》1989,30(4):337-342
Summary A convenient and efficient method of NADPH production from NADP+ has been established using a glucose dehydrogenase system involving whole cells and immobilized cells of Gluconobacter suboxydans IFO 3172. Using airdried cells of the bacterium, the optimum conditions for NADPH production were examined, including the cell and glucose concentrations, NADP+ concentration, pH, buffer and reaction temperature. Under suitable conditions, the conversion ratio and the amount of NADPH accumulated reached about 100% and 73 mg/ml of the reaction mixture, respectively, after 1-h reaction. Intact cells of the bacterium also showed high NADPH production even in the reaction mixture without a surfactant. The addition of Triton X-100 to the reaction mixture and freeze-thawing treatment of intact cells enhanced the production. The NADPH production method was further improved by using cells of the bacterium immobilized by entrapment in a -carrageenan gel lattice. The immobilized cells had almost the same enzymatic properties as the air-dried cells. The conditions for the continuous production of NADPH with an immobilized cell column were also investigated. NADPH was produced in a good yield (about 95%) with this continuous process. 相似文献
5.
Toshikuni Sasaoka Norio Kaneda Yoshikazu Kurosawa Keisuke Fujita Toshiharu Nagatsu 《Neurochemistry international》1989,15(4):555-565
The chromosomal gene for human phenylethanolamine N-methyltransferase (PNMT; EC 2.1.1.28) was isolated from a human genomic library using a cloned human PNMT cDNA as a probe, and the nucleotide sequence was determined. PNMT is encoded in a single gene which consists of three exons. We observed newly the presence of minor PNMT mRNA (type B) besides the major mRNA (type A) as reported previously (Kaneda et al., J. Biol. Chem. 263, 7672–7677, 1988) by Northern hybridization. Type B mRNA carries an approximately 700 nucleotide-long untranslated region in the 5′ terminus. This suggests that two types of mRNA are produced from a single gene through the use of two alternative promoters. A TATA-like sequence locates 30 base pair upstream from the cap site of type A mRNA. Upstream of the cap site, there are several sequences resembling Spl binding sites and glucocorticoid responsive elements, with the latter also found in the first intron. 相似文献
6.
M Numazawa A Mutsumi K Hoshi R Koike 《Biochemical and biophysical research communications》1989,160(3):1009-1014
3-Deoxy steroids having a 4-ene system were found to be competitive inhibitors of human placental aromatase. 19-Hydroxy-4-androsten-17-one (2) potently inhibits the enzyme with an apparent Ki of 12.5 nM, but does not produce a time-dependent inactivation of the enzyme. 相似文献
7.
Mitoshi Yokota Yutaka Tagawa Daikichi Okada Tooru Yasutake Yoshikazu Mine Hiroshi Ishikawa Kousei Miyashita Masao Tomita Sumihiro Tabuchi 《Biotherapy》1990,2(3):207-212
Pre- and postoperative intradermal administration of OK-432 enhanced the SU-PS skin reaction in patients with gastric cancer, but failed to prevent a fall in the NK activity induced by the operation.The change in NK activity was not associated with a change in the proportion of Leu 7-positive cells, but was related to Leu 11a-positive cells. Intradermal injection of OK-432 increased the proportion of Leu 7-positive cells in the patients in whom they accounted for less than 20% of lymphocyte population. The case was the same with Leu 11a-positive cells.Intravenous injection of OK-432 tended to increase suppressor-inducer T cells (CD4+2HA+ cells), B cells and Leu 7-positive cells. Particularly, the proportions of OK-M1-positive cells and MHC class II antigen-positive cells increased in all patients. Immunotherapy with OK-432 given intravenously at a dose of 0.1 KE appeared to be safe because no side effects were essentially observed. 相似文献
8.
Voltage-dependent K+ currents and underlying single K+ channels in pheochromocytoma cells 总被引:11,自引:4,他引:7 下载免费PDF全文
Properties of the whole-cell K+ currents and voltage-dependent activation and inactivation properties of single K+ channels in clonal pheochromocytoma (PC-12) cells were studied using the patch-clamp recording technique. Depolarizing pulses elicited slowly inactivating whole-cell K+ currents, which were blocked by external application of tetraethylammonium+, 4-aminopyridine, and quinidine. The amplitudes and time courses of these K+ currents were largely independent of the prepulse voltage. Although pharmacological agents and manipulation of the voltage-clamp pulse protocol failed to reveal any additional separable whole-cell currents in a majority of the cells examined, single-channel recordings showed that, in addition to the large Ca++-dependent K+ channels described previously in many other preparations, PC-12 cells had at least four distinct types of K+ channels activated by depolarization. These four types of K+ channels differed in the open-channel current-voltage relation, time course of activation and inactivation, and voltage dependence of activation and inactivation. These K+ channels were designated the Kw, Kz, Ky, and Kx channels. The typical chord conductances of these channels were 18, 12, 7, and 7 pS in the excised configuration using Na+-free saline solutions. These four types of K+ channels opened in the presence of low concentrations of internal Ca++ (1 nM). Their voltage-dependent gating properties can account for the properties of the whole-cell K+ currents in PC-12 cells. 相似文献
9.
Thermoluminescence dosimetry measurements of gamma rays produced by the atomic bomb in Hiroshima were made by the predose technique using eight ceramic samples collected from five buildings located at distances between 1271 and 2051 m from the hypocenter. The results of our measurements are compared to both the newer dose estimates (Dosimetry System 1986) and older dose estimates (Tentative 1965 Doses) for survivors of the Hiroshima atomic bomb. In comparison with the older estimates, our results are larger by a factor of 2.3 at 1271 m and 3.9 at 2051 m. Our results and the newer estimates for Hiroshima differ by a factor of only 1.14 +/- 0.16 on the average. 相似文献
10.
Comparative study on the distribution of six species of bitterling in a creek in Fukuoka Prefecture was carried out. The upper part of the creek was fluvial, and the lower part stagnant.Acheilognathus lanceolatus, A. tabira subsp. (b) of Nakamura (1969),A. rhombeus andRhodeus atremius showed wide distribution in the creek throughout their life.A. limbatus seemed to be a fluvial species, andR. ocellatus smithii a lentic species. In the spawning season, however, all the species of bitterling, exceptingR. o. smithii, probably possess a common spawning ground in which mussels occur at a relatively high density. Adult females ofA. lanceolatus, A. tabira subsp. (b) andR. atremius without ripe eggs aggregated in the lower, stagnant parts of the creek. Most juveniles of these species and ofA. rhombeus seemed to grow in the lower part of the creek, and then migrate upward untill their spawning season. No species among the same phylogenetic group has the same set of the life modes (spawning season, spawning sites, aggregation of females without ripe eggs in the lower part of the creek and probability of upstream migration of 0-year-old fish toward spawning sites). 相似文献