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11.
Catalytically competent human and bovine zeta-thrombin and chimeras generated from unfolded polypeptide chains.
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S. D. Lewis D. V. Brezniak J. W. Fenton nd J. A. Shafer 《Protein science : a publication of the Protein Society》1992,1(8):998-1006
Human and bovine alpha-thrombin cleaved at the B-chain by chymotrypsin generates catalytically competent zeta-thrombins, which are comprised of two noncovalently linked fragments: a 36-(human) or 49-(bovine) residue A-chain linked by a disulfide to B-chain residues B1-148 (zeta 1-thrombin) and B-chain residues B149-259 (zeta 2-thrombin). Human and bovine D-Phe-Pro-Arg-CH2-zeta- and PhMeSO2-zeta-thrombins were prepared by reaction of the active-site histidine (H-B43) and serine (S-B205) with PPACK and PMSF, respectively. Unfolding and dissociation of the noncovalently linked polypeptide chains of either human or bovine D-Phe-Pro-Arg-CH2-zeta- and PhMeSO2-zeta-thrombins in 4.5 M guanidine-HCl and refolding upon 30-fold dilution in 50 mM sodium phosphate buffer pH 6.5, 750 mM NaCl, 0.1% PEG resulted in biphasic generation of catalytic activity. The slow phase was eliminated in the presence of the competitive inhibitor benzamidine-HCl. Unfolding and refolding mixtures of the appropriate inactive precursors generated the active chimeric thrombins bovine zeta 1-thrombin:human zeta 2-thrombin and human zeta 1-thrombin:bovine zeta 2-thrombin. Human zeta 1-thrombin and zeta 2-thrombin were isolated, and, upon recombining, the isolated fragments refolded to generate catalytically competent zeta-thrombin with an active-site content, specific activity toward Chromozym-TH, and a specificity constant (kcat/Km) for FPA release from fibrinogen that were all within 60% of those of native alpha-thrombin.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
12.
Concentrations of manganese were determined in the liver, kidney, muscle and bone of white suckers (Catostomus commersoni) from five acid (pH < 5.8), and two circumneutral lakes in south-central Ontario. Manganese tissue concentrations were greater in fish captured from the most acidified lakes with the greatest concentrations of dissolved manganese. These fish had increased concentrations of manganese in the liver, as indicated by a comparison of liver:kidney manganese concentration ratios among the seven fish populations. Tissue concentrations of manganese from all populations either were negatively correlated (P < 0.05) or remained constant with fish size indicating homeostatic regulation of this metal. Manganese concentrations of the benthic fauna were positively correlated to sediment concentrations (R=0.30). Lake sediment manganese concentrations were significantly correlated to maximum lake depth (R=0.80, P < 0.03), with the concentrations in the top 0–1 cm dependent on the redox conditions in the seven lakes. Based on the seven lakes studied, manganese concentrations in the benthic-feeding white sucker correlated better with dissolved manganese, than with either the concentrations in food or surficial sediments. 相似文献
13.
P release rates in five lakes were estimated from a mass balance approach and from laboratory tests. The influence of various environmental parameters on P release has been considered. The internal P loading can contribute phosphorus to overlying waters at levels comparable to the external sources (from 30 to 100%). When considering the recovery from eutrophication of the investigated lakes, the nutrient release from sediments is of critical importance. 相似文献
14.
A vessel was constructed for growth of photosynthetic bacteria at defined light intensity, temperature and partial pressure of oxygen.Under growth conditions at light intensities larger than 1,000 lx, the particles exposed by freeze-fracturing of thylakoids are unordered.Under growth conditions at light intensities lower than 30 lx, the particles seen are hexagonally arranged. If the oxygen partial pressure is increased from 0 to 30 mm Hg while keeping the light intensity at 30 lx, the particles seen in the thylakoids are found to be unregularly arranged.The protein pattern of thylakoids isolated from bacteria grown either at 2,000 lx or at 30 lx revealed a constant ratio of reaction centre polypeptide to either of the membrane polypeptides of 8 kdalton apparent Mr and 12 kdalton apparent Mr.Dedicated to Prof. Dr. G. Drews on occasion of his 60th birthday 相似文献
15.
Effect of mycorrhizal inocula on the growth of Sitka spruce seedlings in different soils 总被引:1,自引:0,他引:1
Summary Different mycorrhizal fungi were tested for their effectiveness in promoting growth of Sitka spruce seedlings, in two contrasting soils, in a glasshouse pot experiment. In nursery soil,Laccaria amethystina significantly improved growth of seedlings in comparison toL. laccata. Seedlings inoculated with a forest isolate ofThelephora terrestris were significantly larger than those inoculated with a nursery isolate when grown in forest soil. The effectiveness ofComplexipes moniliformis in forest soil was poor in comparison to other mycorrhizal fungi. Strains aswell as species of mycorrhizal fungi affect seedling growth differently. These effects are further influenced by soil type. 相似文献
16.
J Ogden 《Journal of medical primatology》1983,12(6):343-345
The reproductive performance of tamarins (Saguinus mystax mystax) under laboratory conditions was evaluated. Fifteen of 20 male-female pairs were fertile and produced 30 live births during an observation period of 3.5 years. Twenty-six breeders (65%) and 12 offspring (40%) more than 3 years old were alive at the termination of the reporting period. 相似文献
17.
A chemiosmotic molecular mechanism for proton-translocating adenosine triphosphatases 总被引:16,自引:0,他引:16
P Mitchell 《FEBS letters》1974,43(2):189-194
18.
Growth versus molting time of caterpillars as a function of temperature,nutrient concentration and the phenolic rutin 总被引:1,自引:0,他引:1
Nancy E. Stamp 《Oecologia》1990,82(1):107-113
Summary A factorial experiment tested the effects of varying nutrient concentration (normal versus diluted), presence or absence of the phenolic allelochemical rutin and daytime temperature (20, 25 and 30° C) on growth, molting and food utilization efficiencies of tobacco hornworms (Manduca sexta). Two of the utilization efficiencies (approximate digestibility and efficiency of conversion of ingested food) were unaffected by temperature; the third one, efficiency of conversion of digested food, was affected by temperature but there was no consistent effect. Lower temperatures significantly increased the proportion of the stadium spent molting, with larvae at a daytime temperature of 20° C spending 9% more of the stadium in molting than larvae at 30° C. Growth time was not influenced by nutrient concentration. When temperature was low and rutin absent, molt time and the proportion of the stadium spent molting were affected by nutrient concentration. Addition of the phenolic rutin did not have an appreciable effect on growth time or digestive processes. However, it increased molting time by 7 to 14% and thus increased the duration of the stadium and reduced relative growth rate. These results indicate that the effect of food quality on growth rate is a function of the thermal conditions of insect herbivores. At cooler temperatures, a disproportionate increase in time spent molting, rather than altered food utilization efficiencies, contributed to lower growth rates. The consequences for larval growth of fluctuating temperatures due to diurnal cycles and the presence of predators forcing larvae into thermally suboptimal microhabitats are discussed. 相似文献
19.
Versieck Jacques Vanballenberghe Lidia Wittoek Ann Vermeir Gerda Vandecasteele Carlo 《Biological trace element research》1990,(1):683-689
A method is described for the determination of mercury in human blood serum and packed blood cells employing neutron activation
analysis. Great attention was devoted to the collection and manipulation of the samples. The accuracy and precision of the
method were tested by analyzing biological reference materials and by comparing the concentrations measured in a number of
serum samples to those obtained by another, independent technique (cold vapor atomic absorption spectrometry) in the same
samples. The article reports the levels measured in blood serum and packed blood cells samples from 15 adult volunteers, as
well as the figures determined in a “second-generation” biological reference material (freeze-dried human serum), prepared
and conditioned at the University of Ghent. 相似文献
20.
Edward J. B. Beeley P. A. Bennett L. G. I. Poland J. S. 《Biological trace element research》1990,(1):53-61
A microcomputer-controlled irradiation and measurement system and a microprocessor-controlled sample changer have been installed
at the SLOWPOKE-2 Facility at the Royal Military College of Canada (RMC). These systems can provide the gamut of instrumental
neutron activation analysis (INAA) techniques for the analyst. Custom software has been created for system control, data acquisition,
and off-line spectral analysis using programs that incorporate Gaussian peak-fitting methods of analysis. The design and use
of the equipment is discussed, and the performance is illustrated with results obtained from the analysis of marine sediment
and biological reference materials. 相似文献