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1.
Properties of purified colchicine-binding protein from a cultured carrot cell extract 总被引:1,自引:0,他引:1
Colchicine-binding protein (CBP) was purified from a cultured carrot cell extract by DEAE-Sephacel, phosphocellulose and Sephadex G200 column chromatographies. The purified CBP separated into three bands on SDS-polyacrylamide gel electrophoresis. One of them reacted with a monoclonal antibody against chick brain alpha-tubulin and the other two with that against beta-tubulin. Colchicine-binding activity of the purified protein was enhanced by tartrate and inhibited little by an excess of podophyllotoxin. It decayed following first order kinetics, but was more stable than the CBP in the crude extract. The binding constant of the purified CBP for colchicine was 0.57 microM-1 and the number of binding sites of colchicine per mg protein was about 2 nmol. This binding constant is about ten times lower than that of porcine brain tubulin under identical conditions. 相似文献
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3.
Kazuo Okamura Kazuyasu Kisaichi Ken-ichiro Takamiya Mitsuo Nishimura 《Archives of microbiology》1984,139(2-3):143-146
A soluble cytochrome, cytochrome c-551 was purified from an aerobic photosynthetic bacterium Erythrobacter species strain OCh 114 (ATCC No. 33942) by ammonium sulfate fractionation, ion-exchange chromatography and gel-filtration. The cytochrome had absorption maxima at 277, 410, and 524–525 nm in the oxidized form, and at 415, 522, and 550.5 nm in the reduced form. At 77 K, the -band of the absorption spectrum of the reduced form split in two at 547 and 549 nm. The millimolar absorption coefficient at 550.5 nm was 26.8 mM-1 cm-1 in the reduced form. This cytochrome was an acidic protein with an isoelectric point of 4.9. Its molecular weight was determined to be 15,000 by gel-filtration on Sephadex G-100 and 14,500 by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The midpoint potential of this cytochrome was +250 mV at pH 7.0. This cytochrome did not bind CO. 相似文献
4.
Synthetic inhibitors of trypsin, plasmin, kallikrein, thrombin, C1r-, and C1 esterase. 总被引:6,自引:0,他引:6
p-Carbethoxyphenyl episol-guanidinocaproate and p-(p'-guanidinobenzoyloxy)-phenyl derivatives were prepared, and their inhibitory effects on trypsin, plasmin, plasma kallikrein, thrombin, C1r- and C1 esterase were examined. Among the various inhibitors tested, p-nitrophenyl p'-guanidinobenzoate, N,N-dimethylamino p-(p'-guanidinobenzoyloxy)-benzoyl glycolate and N,N-dimethylamino p-(p'-guanidinobenzoyloxy)-benzilcarbonyloxy glycolate were the most effective inhibitors of trypsin, plasmin, plasma kallikrien and thrombin, and they strongly inhibited the esterolytic activities of C1r- and C1 esterase. 相似文献
5.
K Okamura Y Sato T Matsuda R Hamanaka M Ono K Kohno M Kuwano 《The Journal of biological chemistry》1991,266(29):19162-19165
Tumor necrosis factor-alpha (TNF-alpha) has been shown to enhance the synthesis of interleukin-6 (IL-6) and collagenase in human omental microvascular endothelial (HOME) cell (Mawatari, M., Kohno, K., Mizoguchi, H., Matsuda, T., Asoh, K., Van Damme, J. V., Welgus, H. G., and Kuwano, M. (1989) J. Immunol. 143, 1619-1627). In the present study, we have examined whether the TNF-alpha-induced synthesis of IL-6 or collagenase in HOME cells is mediated by an inducible growth factor. Among several growth factors examined, we found that the expression of basic fibroblast growth factor (bFGF) mRNA was the one most prominently enhanced when HOME cells were treated with TNF-alpha. The increase of bFGF mRNA by TNF-alpha in HOME cells was observed in both a dose- and time-dependent manner when assayed by Northern blot analysis. The induction of bFGF mRNA was observed by 3 h after incubation with TNF-alpha, and the maximal increase of 5-fold was obtained after 12 h of incubation with 100 units/ml TNF-alpha. Western blot analysis confirmed the enhanced synthesis of bFGF by TNF-alpha. Metabolic labeling and immunoprecipitation assays of bFGF showed that exposure to TNF-alpha enhanced secretion of bFGF into culture medium and also that TNF-alpha stimulated the production of bFGF molecules with molecular masses of 18, 21, and 22.5 kDa in HOME cells. TNF-alpha induced the expression of collagenase mRNA and IL-6 mRNA in HOME cells as well, and the coadministration of neutralizing anti-bFGF antibody almost completely blocked the effects of TNF-alpha. The treatment of HOME cells with exogenous bFGF significantly stimulated the expression of collagenase mRNA and IL-6 mRNA in HOME cells. Therefore, the biological effects of TNF-alpha on HOME cells may be mediated, at least in part, by TNF-alpha-induced bFGF. 相似文献
6.
Possible role of nitric oxide in transmitting information from vasodilator nerve to cerebroarterial muscle 总被引:5,自引:0,他引:5
Treatment with L-NG-monomethyl arginine (L-NMMA), an inhibitor of nitric oxide (NO) synthesis from L-arginine, suppressed the relaxant response of dog cerebral artery strips to transmural electrical stimulation and nicotine, as did oxyhemoglobin. The inhibition by L-NMMA was reversed or prevented by L-, but not D-, arginine. It is concluded that NO or an NO-related compound may play a crucial role in transmitting information from excited vasodilator nerves to cerebroarterial smooth muscle. 相似文献
7.
Effects of chitin/chitosan and their oligomers/monomers on release of type I collagenase from fibroblasts 总被引:2,自引:0,他引:2
The effects of chitin/chitosan and their oligomers/monomers on the release of type I collagenase (MMP-1) from fibroblasts were evaluated using adult (adFB) and neonatal human fibroblasts (neFB) by a immunological assay. Release of MMP-1 from adFB increased significantly or tended to increase for all samples, while there was no significant change in MMP-1 levels with neFB. Because the oligomers and monomers of chitin and chitosan influenced MMP-1 activity, it was suggested that the elevated MMP-1 activity would continue until biodegradation of chitin and chitosan was complete. 相似文献
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9.
Diurnal changes in needle gas exchange in alpine Pinus pumila during snow-melting and summer seasons
Atsushi Ishida Takashi Nakano Seikoh Sekikawa Emiko Maruta Takehiro Masuzawa 《Ecological Research》2001,16(1):107-116
Pinus pumila (Pallas) Regel. is a dominant dwarf tree in alpine regions of Japan. The possible factors limiting the net photosynthetic rate (Pn) of the needles of P. pumila were examined in the snow-melting (May) and the summer (August) seasons. In August, in situ maximum Pn was 20 mol kg–1 needle s–1 in the current-year needles and 25 mol kg–1 needle s–1 in the 1-year-old needles. Diurnal trends of Pn in August were positively related to fluctuations in photosynthetic photon flux density (PPFD) and no midday depression of Pn was found, indicating that a decrease in PPFD rather than an increase in needle-to-air vapor pressure deficit (W) might cause the reduction of Pn. Both stomatal conductance (gs) and Pn were lower in May than in August. In May, Pn and gs were almost zero in the morning, but gradually increased with decreasing W, reaching maximum Pn values (4 mol kg–1 needle s–1) and gs (60 mmol kg–1 needle s–1) at 16.00 hours. The daytime Pn in May was positively related to gs. Relative water content in the exposed needles above the snow in May was 83%, which was far above the lethal level. This indicates that the water flow from stems or soils to needles was enough to compensate for a small amount of water loss due to the low gs in May, although the water supplied to needles would be impeded by the low temperatures. Thus, the reduced gs in May would be important for avoiding needle desiccation, and would result in a reduced Pn. 相似文献
10.
Mizukami Y Okamura T Miura T Kimura M Mogami K Todoroki-Ikeda N Kobayashi S Matsuzaki M 《Biochimica et biophysica acta》2001,1540(3):213-220
Cytokines and various cellular stresses are known to activate c-Jun N-terminal kinase-1 (JNK1), which is involved in physiological function. Here, we investigate the activation of JNK1 by oxidative stress in H9c2 cells derived from rat cardiomyocytes. H(2)O(2) (100 microM) significantly induces the tyrosine phosphorylation of JNK1 with a peak 25 min after the stimulation. The amount of JNK1 protein remains almost constant during stimulation. Immunocytochemical observation shows that JNK1 staining in the nucleus is enhanced after H(2)O(2) stimulation. To clarify the physiological role of JNK1 activation under these conditions, we transfected antisense JNK1 DNA into H9c2 cells. The antisense DNA (2 microM) inhibits JNK1 expression by 80% as compared with expression in the presence of the sense DNA, and significantly blocks H(2)O(2)-induced cell death. Consistent with the decrease in cell number, we detected condensation of the nuclei, a hallmark of apoptosis, 3 h after H(2)O(2) stimulation in the presence of the sense DNA for JNK1. The antisense DNA of JNK1 inhibits the condensation of nuclei by H(2)O(2). Under these conditions, the H(2)O(2)-induced phosphorylation of proteins with molecular masses of 55, 72, and 78 kDa is blocked by treatment with the antisense DNA for JNK1 as compared with the sense DNA for JNK1. These findings suggest that JNK1 induces apoptotic cell death in response to H(2)O(2), and that the cell death may be involved in the phosphorylations of 55, 72, and 78 kDa proteins induced by JNK1 activation. 相似文献