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31.
Kimler VA Palazzolo KL Anne P Haddad MM Lee JB Harkins C Vallarapu B Taylor JD 《Pigment cell research / sponsored by the European Society for Pigment Cell Research and the International Pigment Cell Society》2002,15(4):298-304
Fish chromatophores serve as excellent study models for cytoskeleton-dependent organelle translocations because the distribution of pigmentary organelles can be observed against a time frame by microscopy. In this study the distribution of microfilaments along with microtubules in cultured melanophores of the killifish (Fundulus heteroclitus Linneaus) are examined using whole-cell transmission electron microscopy (WCTEM), fluorescence, and laser scanning confocal microscopy. Dispersing, dispersed, aggregating and aggregated states of pigment are induced by adding either caffeine (for dispersion) or epinephrine (for aggregation) to the cells in a standard culture medium. The cells that exhibited a random melanosome distribution in the standard culture media without these two reagents, served as the control. The results indicate that: (i) a structure considered to be the actin-filament organizing center (AFOC) is in close proximity to the microtubule-organizing center (MTOC); (ii) the radial layout of microfilaments remains similar over four physiological states of pigmentary response with the exception of epinephrine-aggregated pigment, in which the aggregate blocks the viewing of the AFOC and central microfilament rays, yet radial microfilaments, whether central and/or peripheral, are apparent in all physiological states of distribution; and (iii) microfilaments serve, together with microtubules, as scaffolding for melanosomes which migrate in bi-directional rows on cross-bridges, thus shedding light on the mechanisms for orderly melanosome translocations in a structural continuum. 相似文献
32.
Bhushan B Trott S Spain JC Halasz A Paquet L Hawari J 《Applied and environmental microbiology》2003,69(3):1347-1351
A unique metabolite with a molecular mass of 119 Da (C(2)H(5)N(3)O(3)) accumulated during biotransformation of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) by Rhodococcus sp. strain DN22 (D. Fournier, A. Halasz, J. C. Spain, P. Fiurasek, and J. Hawari, Appl. Environ. Microbiol. 68:166-172, 2002). The structure of the molecule and the reactions that led to its synthesis were not known. In the present study, we produced and purified the unknown metabolite by biotransformation of RDX with Rhodococcus sp. strain DN22 and identified the molecule as 4-nitro-2,4-diazabutanal using nuclear magnetic resonance and elemental analyses. Furthermore, we tested the hypothesis that a cytochrome P450 enzyme was responsible for RDX biotransformation by strain DN22. A cytochrome P450 2B4 from rabbit liver catalyzed a very similar biotransformation of RDX to 4-nitro-2,4-diazabutanal. Both the cytochrome P450 2B4 and intact cells of Rhodococcus sp. strain DN22 catalyzed the release of two nitrite ions from each reacted RDX molecule. A comparative study of cytochrome P450 2B4 and Rhodococcus sp. strain DN22 revealed substantial similarities in the product distribution and inhibition by cytochrome P450 inhibitors. The experimental evidence led us to propose that cytochrome P450 2B4 can catalyze two single electron transfers to RDX, thereby causing double denitration, which leads to spontaneous hydrolytic ring cleavage and decomposition to produce 4-nitro-2,4-diazabutanal. Our results provide strong evidence that a cytochrome P450 enzyme is the key enzyme responsible for RDX biotransformation by Rhodococcus sp. strain DN22. 相似文献
33.
The microbial and enzymatic degradation of a new energetic compound, 2,4,6,8,10,12-hexanitro-2,4,6,8,10,12-hexaazaisowurtzitane (CL-20), is not well understood. Fundamental knowledge about the mechanism of microbial degradation of CL-20 is essential to allow the prediction of its fate in the environment. In the present study, a CL-20-degrading denitrifying strain capable of utilizing CL-20 as the sole nitrogen source, Pseudomonas sp. strain FA1, was isolated from a garden soil. Studies with intact cells showed that aerobic conditions were required for bacterial growth and that anaerobic conditions enhanced CL-20 biotransformation. An enzyme(s) involved in the initial biotransformation of CL-20 was shown to be membrane associated and NADH dependent, and its expression was up-regulated about 2.2-fold in CL-20-induced cells. The rates of CL-20 biotransformation by the resting cells and the membrane-enzyme preparation were 3.2 +/- 0.1 nmol h(-1) mg of cell biomass(-1) and 11.5 +/- 0.4 nmol h(-1) mg of protein(-1), respectively, under anaerobic conditions. In the membrane-enzyme-catalyzed reactions, 2.3 nitrite ions (NO(2)(-)), 1.5 molecules of nitrous oxide (N(2)O), and 1.7 molecules of formic acid (HCOOH) were produced per reacted CL-20 molecule. The membrane-enzyme preparation reduced nitrite to nitrous oxide under anaerobic conditions. A comparative study of native enzymes, deflavoenzymes, and a reconstituted enzyme(s) and their subsequent inhibition by diphenyliodonium revealed that biotransformation of CL-20 is catalyzed by a membrane-associated flavoenzyme. The latter catalyzed an oxygen-sensitive one-electron transfer reaction that caused initial N denitration of CL-20. 相似文献
34.
Effect of amino acids on production of xylanase and pectinase from Streptomyces sp. QG-11-3 总被引:5,自引:0,他引:5
Qasim Khalil Beg Bharat Bhushan Mukesh Kapoor G.S. Hoondal 《World journal of microbiology & biotechnology》2000,16(2):211-213
Xylanase and pectinase production by Streptomyces sp. QG-11-3 was stimulated by DL-norleucine, L-leucine, DL-isoleucine, L-lysine monohydrochloride and DL--phenylalanine by up to 3.72- and 2.78-fold, respectively, whereas the combination of DL-norleucine, L-leucine and DL-isoleucine synergistically stimulated the xylanase and pectinase production by up to 6.72- and 5.62-fold, respectively. Glycine, DL-norvaline, DL-methionine, and DL-aspartic acid showed no significant stimulatory effect on enzyme production. 相似文献
35.
Potentially, silver development could unify most modern demands for clean, accurately localized marker amplification in microscopy and bioanalysis. However, the existing technology leaves room for improvement in developer design. A new formulation has been devised which, by using principles of silver chelation, avoids problems of self-nucleation and catalysis by light. It is made, just before use, by mixing together equal amounts of stock solutions containing high molarity, Tris-buffered silver nitrate and alcoholic, buffered pyrogallol. The two stocks are easily prepared and have very long shelf-lives. The developer is light insensitive for up to an hour at room temperature, so that development can proceed under ambient light conditions and at the neutral pH most suited to biological systems. The powerful reducer in the suggested formulation should allow the detection of low concentrations of marker signal in a wide range of applications. 相似文献
36.
An alkalophilic, chitinase-producing Bacillus sp. BG-11 was isolated which produced an extracellular chitinase and which was purified 16.5-fold, using standard purification techniques. The purified chitinase exhibited a broad pH and temperature optima of 7.5-9.0 and 45 deg C-55 deg C, respectively. The chitinase was stable between pH 6.0-9.0 and 50°C for more than 2 h. Half lives of enzyme at 60 deg C, 70 deg C and 80 deg C were 90 min, 30 min and 20 min respectively. Km value was 12 mg chitin per ml. Shelf life was 60 days at 4°C. Ca2+, Ni2+ and Triton-X-100 stimulated the activity up to 20% whereas Ag+, Hg2+, dithiothreitol, -mercaptoethanol, glutathione, iodoacetic acid and iodoacetamide inhibited the activity up to 50%. 相似文献
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Mitogen-activated protein kinase kinase-4 promotes cell survival by decreasing PTEN expression through an NF kappa B-dependent pathway 总被引:2,自引:0,他引:2
Xia D Srinivas H Ahn YH Sethi G Sheng X Yung WK Xia Q Chiao PJ Kim H Brown PH Wistuba II Aggarwal BB Kurie JM 《The Journal of biological chemistry》2007,282(6):3507-3519