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1.
Antizyme inhibitor was highly purified from rat liver by using affinity chromatography. It has some structural resemblance to ornithine decarboxylase (ODC), as judged from Mr, immunoreactivity and reversible binding with antizyme. However, unlike hepatic amounts of ODC and ODC-antizyme complex, that of antizyme inhibitor did not show much fluctuation upon putrescine treatment, whereas it decreased as rapidly as ODC decay in the presence of cycloheximide. These results suggested that antizyme inhibitor is an independent regulatory protein rather than a derivative of ODC. Changes in hepatic amounts of antizyme inhibitor, antizyme and ODC upon feeding suggested that antizyme inhibitor may play a role in ODC regulation by trapping antizyme and thereby suppressing ODC degradation. A monoclonal antibody to rat liver antizyme inhibitor was obtained. This antibody was shown to be utilizable for a simple assay of antizyme-inhibitor activity in tissue extracts. 相似文献
2.
Antizyme to ornithine decarboxylase is present in the liver of starved rats. 总被引:1,自引:1,他引:0
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Antizyme to ornithine decarboxylase (ODC) and ODC-antizyme complex were both present in liver cytosols of starved rats. The antizyme was identified by its molecular weight, kinetic properties, formation of a complex with ODC, and reversal of its inhibition by antizyme inhibitor. The average amount of antizyme in liver cytosols of starved rats was 0.1 unit/mg of protein, roughly corresponding to basal hepatic ODC activity in rats fed ad libitum. The presence of ODC-antizyme complex was detected by using antizyme inhibitor. These results indicate that antizyme participates in the regulation of ODC activity in vivo under physiological conditions. 相似文献
3.
Isawa H Orito Y Jingushi N Iwanaga S Morita A Chinzei Y Yuda M 《The FEBS journal》2007,274(16):4271-4286
Two plasma kallikrein-kinin system inhibitors in the salivary glands of the kissing bug Triatoma infestans, designated triafestin-1 and triafestin-2, have been identified and characterized. Reconstitution experiments showed that triafestin-1 and triafestin-2 inhibit the activation of the kallikrein-kinin system by inhibiting the reciprocal activation of factor XII and prekallikrein, and subsequent release of bradykinin. Binding analyses showed that triafestin-1 and triafestin-2 specifically interact with factor XII and high molecular weight kininogen in a Zn2+-dependent manner, suggesting that they specifically recognize Zn2+-induced conformational changes in factor XII and high molecular weight kininogen. Triafestin-1 and triafestin-2 also inhibit factor XII and high molecular weight kininogen binding to negatively charged surfaces. Furthermore, they interact with both the N-terminus of factor XII and domain D5 of high molecular weight kininogen, which are the binding domains for biological activating surfaces. These results suggest that triafestin-1 and triafestin-2 inhibit activation of the kallikrein-kinin system by interfering with the association of factor XII and high molecular weight kininogen with biological activating surfaces, resulting in the inhibition of bradykinin release in an animal host during insect blood-feeding. 相似文献
4.
Chattopadhyay MK Murakami Y Matsufuji S 《The Journal of biological chemistry》2001,276(24):21235-21241
The mechanism of the regulatory degradation of ornithine decarboxylase (ODC) by polyamines was studied in fission yeast, Schizosaccharomyces pombe. To regulate cellular spermidine experimentally, we cloned and disrupted S-adenosylmethionine decarboxylase gene (spe2) in S. pombe. The null mutant of spe2 was devoid of spermidine and spermine, accumulated putrescine, and contained a high level of ODC. Addition of spermidine to the culture medium resulted in rapid decrease in the ODC activity caused by the acceleration of ODC degradation, which was dependent on de novo protein synthesis. A fraction of ODC forming an inactive complex concomitantly increased. The accelerated ODC degradation was prevented either by knockout of antizyme gene or by selective inhibitors of proteasome. Thus, unlike budding yeast, mammalian type antizyme-mediated ODC degradation by proteasome is operating in S. pombe. 相似文献
5.
Olga?V?Matveeva Brian?T?Foley Vladimir?A?Nemtsov Raymond?F?Gesteland Senya?Matsufuji John?F?Atkins Aleksey?Y?Ogurtsov Svetlana?A?ShabalinaEmail author 《BMC bioinformatics》2004,5(1):44
Background
Computer programs for the generation of multiple sequence alignments such as "Clustal W" allow detection of regions that are most conserved among many sequence variants. However, even for regions that are equally conserved, their potential utility as hybridization targets varies. Mismatches in sequence variants are more disruptive in some duplexes than in others. Additionally, the propensity for self-interactions amongst oligonucleotides targeting conserved regions differs and the structure of target regions themselves can also influence hybridization efficiency. There is a need to develop software that will employ thermodynamic selection criteria for finding optimal hybridization targets in related sequences. 相似文献6.
Antizyme frameshifting as a functional probe of eukaryotic translational termination 总被引:1,自引:1,他引:0
Karamysheva ZN Karamyshev AL Ito K Yokogawa T Nishikawa K Nakamura Y Matsufuji S 《Nucleic acids research》2003,31(20):5949-5956
Translation termination in eukaryotes is mediated by the release factors eRF1 and eRF3, but mechanisms of the interplay between these factors are not fully understood, due partly to the difficulty of measuring termination on eukaryotic mRNAs. Here, we describe an in vitro system for the assay of termination using competition with programmed frameshifting at the recoding signal of mammalian antizyme. The efficiency of antizyme frameshifting in rabbit reticulocyte lysates was reduced by addition of recombinant rabbit eRF1 and eRF3 in a synergistic manner. Addition of suppressor tRNA to this assay system revealed competition with a third event, stop codon readthrough. Using these assays, we demonstrated that an eRF3 mutation at the GTPase domain repressed termination in a dominant negative fashion probably by binding to eRF1. The effect of the release factors and the suppressor tRNA showed that the stop codon at the antizyme frameshift site is relatively inefficient compared to either the natural termination signals at the end of protein coding sequences or the readthrough signal from a plant virus. The system affords a convenient assay for release factor activity and has provided some novel views of the mechanism of antizyme frameshifting. 相似文献
7.
Identification of nuclear export signals in antizyme-1 总被引:8,自引:0,他引:8
Antizyme-1 (AZ1) is a protein that negatively regulates polyamine synthesis by inhibiting the key synthetic enzyme ornithine decarboxylase and targeting it for degradation by the 26 S proteasome. Recent work shows that antizyme protein translocates to the nucleus during mouse development (Gritli-Linde, A., Nilssom, J., Bohlooly, Y. M., Heby, O., and Linde, A. (2001) Dev. Dyn. 220, 259-275). However, the significance and mechanism of this phenomenon remain unclear. In this study, we expressed AZ1 fused with enhanced green fluorescent protein (EGFP) to study its localization in a living cell. We found that EGFP-AZ1 was predominantly localized in the cytoplasm and that treatment with leptomycin B, a specific inhibitor of chromosomal region maintenance 1 (CRM1) induced nuclear accumulation of EGFP-AZ1 in Chinese hamster ovary and NIH3T3 cells. Two independent nuclear export signal (NES) sequences, each containing essential hydrophobic residues, were identified in the 50 N-terminal amino acid residues and in the central part of AZ1. The activity of the second NES was inhibited by an N-terminal adjacent region and was only revealed in N-terminal truncated constructs. Both NESs were active when fused to an artificial nuclear protein SV40-NLS-EGFP-EGFP. The ability of AZ1 to shuttle between the nucleus and the cytoplasm suggests that it has a novel function in the nucleus. 相似文献
8.
Suhara H Daikoku C Takata H Suzuki S Matsufuji Y Sakai K Kondo R 《Applied microbiology and biotechnology》2003,62(5-6):601-607
Bioremediation is a low-cost treatment alternative for the cleanup of polychlorinated-dioxin-contaminated soils and fly ash when pollution spread is wide-ranging. An interesting fungus, Ceriporia sp. MZ-340, with a high ability to degrade dioxin, was isolated from white rotten wood of a broadleaf tree from Kyushu Island in Japan. We have attempted to use the fungus for bioremediation of polychlorinated-dioxin-contaminated soil on site. However, we have to consider that this trial has the potential problem of introducing a biohazard to a natural ecosystem if this organism is naturalized. We have therefore developed a monitoring system for the introduced fungus as a part of the examination and evaluation of bioremediation in our laboratory. We have also developed a PCR-based assay to reliably detect the fungus at the bioremediation site. DNA isolated from the site was amplified by PCR using a specific primer derived from internal transcribed spacer region (ITS: ITS1, 5.8S rDNA and ITS2) sequences of Ceriporia sp. MZ-340. We successfully monitored Ceriporia sp. MZ-340 down to 100 fg/µl DNA and down to 2 mg/g mycelium. We also successfully monitored the fungus specifically at the bioremediation site. The polychlorinated dibenzo-p-dioxin and polychlorinated dibenzofuran content was observed to decrease in response to treatment with the fungus. The species-specific PCR technique developed in the present work is useful in evaluating the possibility of on-site bioremediation using the fungus Ceriporia sp. MZ-340. 相似文献
9.
Hori N Newton RU Andrews WA Kawamori N McGuigan MR Nosaka K 《Journal of strength and conditioning research / National Strength & Conditioning Association》2007,21(2):314-320
Measurement of power output during resistance training is becoming ubiquitous in strength and conditioning programs, but there is great variation in the methods used. The main purposes of this study were to compare the power output values obtained from 4 different methods and to examine the relationships between these values. Male semiprofessional Australian rules football players (n = 30) performed hang power clean and weighted jump squat while ground reaction force (GRF)-time data and barbell displacement-time data were sampled simultaneously using a force platform and a linear position transducer attached to the barbell. Peak and mean power applied to the barbell was obtained from barbell displacement-time data (method 1). Peak and mean power applied to the system (barbell + lifter) was obtained from 3 other methods: (a) using GRF-time data (method 2), (b) using barbell displacement-time data (method 3), and (c) using both barbell displacement-time data and GRF-time data (method 4). The peak power values (W) obtained from methods 1, 2, 3, and 4 were (mean +/- SD) 1,644 +/- 295, 3,079 +/- 638, 3,821 +/- 917, and 4,017 +/- 833 in hang power clean and 1,184 +/- 115, 3,866 +/- 451, 3,567 +/- 494, and 4,427 +/- 557 in weighted jump squat. There were significant differences between power output values obtained from method 1 vs. methods 2, 3, and 4, as well as method 2 vs. methods 3 and 4. The power output applied to the barbell and that applied to the system was significantly correlated (r = 0.65-0.81). As a practical application, it is important to understand the characteristics of each method and consider how power output should be measured during the hang power clean and the weighted jump squat. 相似文献
10.
Matsuda T Fujimura S Suda H Matsufuji Y Nakagawa J 《Bioscience, biotechnology, and biochemistry》2011,75(9):1829-1831
Upon exposure to 8% ethanol, survival and growth of yeast strains deficient in histone deacetylase complex genes was examined. Of the 18 mutants tested, the Δsir3 and Δsir4 strains showed higher resistance to ethanol, while the Δrco1, Δhos3, Δhda2, and Δhst1 strains were more sensitive than the wild type. Furthermore, these ethanol-resistant patterns varied under aerobic and anaerobic culture conditions. 相似文献