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91.
Z Wasylewski H Ko?oczek A Wa?niowska K Slizowska 《European journal of biochemistry》1992,206(1):235-242
The dependence of the fluorescence emission maximum of the tryptophan residues in several two-tryptophan-containing proteins (horse liver alcohol dehydrogenase, yeast 3-phosphoglycerate kinase, Staphylococcus aureus metalloprotease and bee venom phospholipase A2) on the excitation wavelengths has been studied. Using fluorescence-resolved spectroscopy, we have dissected the contributions of particular tryptophan residues located in different parts of the protein molecule. The results demonstrate that dipolar structural relaxation can occur in the environment of tryptophan residues buried within protein molecules. The observed spectral shifts upon red-edge excitation of these residues can depend on temperature or ligand binding, as demonstrated in case of metalloprotease and alcohol dehydrogenase. No spectral shifts upon red-edge excitation have been observed for tryptophan residues totally exposed to the rapidly relaxing aqueous solvent. 相似文献
92.
Eiji Hara Tomoko Ohshima Takako Ishii Wataru Sugino Ko Tsutsui Susumu Nakada Nobuo Tsuchida Kinichiro Oda 《Experimental cell research》1992,198(2):250-258
The mechanism of induction of DNA synthesis in quiescent rat 3Y1 cells by the adenovirus E1A gene was investigated using the 3Y1 derivative cell lines g12-21, gn12RB1, and gn12RB2. The g12-21 cells express the E1A 12S cDNA and the latter two cells express both the E1A 12S cDNA and the human retinoblastoma susceptibility (Rb) gene at different levels in response to dexamethasone (dex). The cDNA sequences of E1A-inducible cell cycle-dependent genes, clone 3 and clone 16, were isolated by differential screening of a cDNA library constructed from dex-treated g12-21 cells. The quiescent 3Y1 cells induced c-fos and c-myc expression within 2 h after serum stimulation and expressed clone 16 and clone 3 transiently at around 8 h before the onset of DNA synthesis (10 h). In contrast, the quiescent g12-21 cells treated with dex expressed a high level of E1A at 6 to 8 h after treatment and expressed clone 16 and clone 3 at around 8 h without stimulation of c-fos and c-myc expression, suggesting that E1A bypasses the cell cycle early in G1. The half-maximal rate of DNA synthesis was reached in a much shorter time in dex-treated g12-21 cells (12 h) than in serum-treated 3Y1 cells (18 h), suggesting that E1A also bypasses the cell cycle at the G1/S boundary. The gn12RB1 and gn12RB2 cells were unable to induce DNA synthesis in response to dex presumably due to lower levels of E1A expression, although gn12RB2 but not gn12RB1 cells could express clone 16 and clone 3. These results suggest that the level of E1A required for bypass at the G1/S boundary is higher than that required early in G1. 相似文献
93.
D K Ann J Hasegawa J L Ko S T Chen N M Lee H H Loh 《The Journal of biological chemistry》1992,267(11):7921-7926
We have recently identified and sequenced the cDNA for an opioid-binding protein with homologies to cell adhesion molecules (OBCAM) (Schofield, P. R., McFarlard, K. C., Hayflick, J. S., Wilcox, J. N., Cho, T. M., Roy, S., Lee, N. M., Loh, H. H., and Seeburg, P. H. (1989) EMBO J. 8, 489-495). Several lines of evidence using antibodies suggest that OBCAM may play a functional role in NG108-15 neuroblastoma x glioma cells, a useful model system that contains a homogeneous population of delta-opioid receptors. A logical extension of this research is to further test this hypothesis. As part of this study, NG108-15 cells were stably transfected with either sense or antisense sequences of a portion of pROM, the rat cDNA for OBCAM. [3H] Diprenorphine binding was greatly reduced in antisense-transfected cells relative to non-transfected cells. Binding to alpha 2-adrenergic, muscarinic, and insulin receptors was unaffected. These results further support the notion that OBCAM or its analogue is part (or a subunit) of an opioid receptor. Furthermore, our observation of an apparently specific reduction in opioid binding in these transfected cells suggests that they may provide a novel genetic approach for studying regulation of the opioid receptor in this defined cell line. 相似文献
94.
Induction of an organ-specific autoimmune disease, lymphocytic hypophysitis, in hamsters by recombinant rubella virus glycoprotein and prevention of disease by neonatal thymectomy. 下载免费PDF全文
J W Yoon D S Choi H C Liang H S Baek I Y Ko H S Jun S Gillam 《Journal of virology》1992,66(2):1210-1214
Glycosylated, membrane-associated E1 (58-kDa) and E2 (47- to 49-kDa) rubella virus proteins and unglycosylated nucleoprotein C (33 kDa), from separately expressed vaccinia virus recombinants, were injected into golden Syrian hamsters. Rubella virus E1 and E2 glycoproteins consistently induced an organ-specific autoimmune disease, autoimmune lymphocytic hypophysitis, which was evidenced by the induction of autoantibodies against pituitary cells and by lymphocytic infiltration of the pituitary. Neonatal thymectomy prevented the disease. In contrast, rubella virus nucleoprotein C did not induce either autoantibodies against pituitary cells or lymphocytic infiltration of the pituitary. This finding raises the possibility that virus-specific protein itself can induce an organ-specific autoimmune disease in certain circumstances. 相似文献
95.
Human sperm karyotypes can be prepared after fusion of human sperm with Golden hamster oocytes. Most laboratories use one of two methods of sperm capacitation: incubation of freshly-ejaculated sperm in Biggers, Whitten, and Whittingham (BWW) medium for 5-7 h at 37 degrees C or sperm storage in (N-tris [hydroxymethyl]methyl-2-aminoethanesulfonic acid; 2-([2-hydroxy-1,1-bis(hydroxymethyl)ethyl]amino)ethanesulfonic acid) (TES)-Tris yolk buffer (TYB) for 1-3 days at 4 degrees C. Since there have been conflicting reports as to whether there is a difference in the frequency of structural chromosomal abnormalities between BWW capacitation and storage in TYB for 2 days, we analyzed a larger number of karyotypes (8974) from 136 donors to determine if there was any difference in the frequency or type of chromosomal abnormalities in sperm treated by fresh BWW capacitation, storage in TYB for 1 day (TYB-1), or storage in TYB for 2 days (TYB-2). There was no difference in the frequency of numerical chromosomal abnormalities or sex ratio in any of the three treatment groups. However, there was a significantly increased frequency of structural chromosomal abnormalities after storage in TYB-1 and TYB-2. There was no difference in the frequency or type of structural chromosomal abnormalities after sperm storage in TYB-1 compared to TYB-2. 相似文献
96.
97.
When oospores from the pairing between A1 and A2 mating types of Phytophthora infestans were treated with 0.25 % KMnO4 solution for 15 min and incubated at 19 °C under light on a modified S+L medium, germination commenced within 4 days and reached about 70 % after 20 days. Under these conditions, more than 25 % of oospores obtained from a 4-day-old culture germinated. To obtain a high germination rate of P. infestans oospores, light was essential during germination but not during growth and oospore formation. The optimum time for activation of oospores with 0.25 % KMnO4 was 15 to 30 min and a suitable concentration of KMnO4 for 15 min activation was 0.25 to 0.45 %. 相似文献
98.
In Saccharomyces cerevisiae yeast, a cycloheximide-sensitive "prestart" step (Ki = 0.12 +/- 0.05 microM) is passaged greater than or equal to 4 to less than or equal to 13 minutes prior to the alpha-factor-sensitive "Start" step in the cell cycle of normal proliferating cells and in the first and second bud emergence cycles of abnormally large cells that had been arrested for cell division with alpha-factor and allowed to recover. This identifies the chronologically last protein synthetic step of the cell cycle that occurs prior to the completion of Start. This step is named the last synthetic prestart or LSP step. Cells require the completion of the LSP step before they can perform Start during recovery from arrest by alpha-factor. Yet alpha-factor is known to prevent cell division by acting at Start. The combined data suggest that alpha-factor prevents the Start step of cell division by inactivating a protein that is 1) required for the performance of Start, and 2) synthesized shortly prior to Start in the last synthetic prestart step. 相似文献
99.
Ferrous Iron and Sulfur Oxidation and Ferric Iron Reduction Activities of Thiobacillus ferrooxidans Are Affected by Growth on Ferrous Iron, Sulfur, or a Sulfide Ore 总被引:4,自引:3,他引:1 下载免费PDF全文
Isamu Suzuki Travis L. Takeuchi Trin D. Yuthasastrakosol Jae Key Oh 《Applied microbiology》1990,56(6):1620-1626
Eight strains of Thiobacillus ferrooxidans (laboratory strains Tf-1 [= ATCC 13661] and Tf-2 [= ATCC 19859] and mine isolates SM-1, SM-2, SM-3, SM-4, SM-5, and SM-8) and three strains of Thiobacillus thiooxidans (laboratory strain Tt [= ATCC 8085] and mine isolates SM-6 and SM-7) were grown on ferrous iron (Fe2+), elemental sulfur (S0), or sulfide ore (Fe, Cu, and Zn). The cells were studied for their aerobic Fe2+ - and S0-oxidizing activities (O2 consumption) and anaerobic S0-oxidizing activity with ferric iron (Fe3+) (Fe2+ formation). Fe2+-grown T. ferrooxidans cells oxidized S0 aerobically at a rate of 2 to 4% of the Fe2+ oxidation rate. The rate of anaerobic S0 oxidation with Fe3+ was equal to the aerobic oxidation rate in SM-1, SM-3, SM-4, and SM-5, but was only one-half or less that in Tf-1, Tf-2, SM-2, and SM-8. Transition from growth on Fe2+ to that on S0 produced cells with relatively undiminished Fe2+ oxidation activities and increased S0 oxidation (both aerobic and anaerobic) activities in Tf-2, SM-4, and SM-5, whereas it produced cells with dramatically reduced Fe2+ oxidation and anaerobic S0 oxidation activities in Tf-1, SM-1, SM-2, SM-3, and SM-8. Growth on ore 1 of metal-leaching Fe2+-grown strains and on ore 2 of all Fe2+-grown strains resulted in very high yields of cells with high Fe2+ and S0 oxidation (both aerobic and anaerobic) activities with similar ratios of various activities. Sulfur-grown Tf-2, SM-1, SM-4, SM-6, SM-7, and SM-8 cultures leached metals from ore 3, and Tf-2 and SM-4 cells recovered showed activity ratios similar to those of other ore-grown cells. It is concluded that all the T. ferrooxidans strains studied have the ability to produce cells with Fe2+ and S0 oxidation and Fe3+ reduction activities, but their levels are influenced by growth substrates and strain differences. 相似文献
100.