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101.
Pétur Reimarsson Björn Lindman Moshe M. Werber 《Archives of biochemistry and biophysics》1980,202(2):664-666
The anion-binding characteristics of ferredoxin from Halobacterium of the Dead Sea have been studied by 35Cl? NMR. It is found that the binding constant of Cl? to halophilic ferredoxin is ca. 0.09 at 28 °C and that the binding enthalpy is positive. It is also found that the correlation time for chloride ions bound to halophilic ferredoxin is about 10 ns. The effect on the 35Cl?1 NMR signal of adding competing anions is also studied. Halophilic proteins like ferredoxin which have a high negative charge bind anions with low affinity but the 35Cl? quadrupole relaxation technique can conveniently monitor such weak binding. 相似文献
102.
Cartilage proteoglycan aggregate formation was studied by zonal rate centrifugation in sucrose gradients. Proteoglycan aggregates, monomers and proteins could be resolved. It was shown that the optimal proportion of hyaluronic acid for proteoglycan aggregate formation was about 1% of proteoglycan dry weight. The reaggregation of dissociated proteoglycan aggregate A1 fraction was markedly concentration-dependent and even at 9 mg/ml only about 90% of the aggregates were reformed. The lowest proportion of link protein required for maximal formation of link-stabilized proteoglycan aggregates was 1.5% of proteoglycan dry weight. It was separately shown that link protein co-sedimented with the proteoglycan monomer. By competition with isolated hyaluronic acid-binding-region fragments, a proportion of the link proteins was removed from the proteoglycan monomers, indicating that the link protein binds to the hyaluronic acid-binding region of the proteoglycan monomer. 相似文献
103.
Summary
Atriplex hymenelytra is an evergreen shrub distributed in the hot deserts of parts of Mexico and the southwestern United States. The leaves of the species have a number of characteristics that are adaptive in a hot, dry environment, some of which change seasonally. Steeply angled leaves reduce midday solar interception, yet result in relatively high interception when solar angles are low and vapor pressure deficits are at a minimum. The leaves substantially reduce their absorptance of incident radiation during the hot periods of the year by changing their moisture and hence dissolved salt contents. At these times the light intensity required for saturation of photosynthesis is low and a reduction in the radiation absorbed by the leaves therefore results in a greater water-use efficiency. 相似文献
104.
P Bj?rntorp M Karlsson H Pertoft P Pettersson L Sj?str?m U Smith 《Journal of lipid research》1978,19(3):316-324
To identify cells developing into adipocytes by accumulation of triglyceride, rat epididymal fat pad cells from small rats were exposed to (3)H-labeled chylomicron fatty acids in vivo and then liberated with collagenase. Tissue remnants were removed by filtration and mature fat cells by flotation. Aggregating cells were then removed by filtration through a 25- micro m nylon screen. Further purification of cells labeled in vivo was obtained by removing floating cells from those adhering to the bottom of a culture dish. The adhering cells multiplied to a confluent monolayer when cultured in Medium 199 containing serum, glucose, insulin, and a triglyceride emulsion. The cells then gradually enlarged due to granulation of the cytoplasm by a lipid-staining material. After about 2 weeks these granules had coalesced forming mature adipocytes of typical signet-ring appearance. Free adipocytes could then be recovered from the cultures by collagenase treatment. After about 2 weeks of culture these cells had the same size (about 30 micro m) as adipocytes recovered in the original collagenase preparation of the rat epididymal fat pad. They contained triglyceride lipase activity and incorporated glucose into triglycerides to the same extent as cells developed in vivo but had higher lipoprotein lipase activity. In vitro, heparin in a low concentration, prostaglandin E(1), isobutylmethylxanthine, and cholera toxin markedly promoted the development of these cells into adipocytes. This could be shown to occur almost completely indicating that this fraction of cells was homogeneous and consisted of cells with the capacity to form adipocytes. The duplication time was about 2 days and did not change with subculturing. Preadipocytes could be obtained by density gradient centrifugation, isolating triglyceride-containing cells either directly from the pad or after 3 days in culture. All of these cells developed into adipocytes as described above but did not multiply as readily. It was concluded that cells from the epididymal fat pad from small rats can be isolated in a homogenous fraction that develops in culture into cells of identical morphology and function as adipocytes formed in vivo. The differentiation of these cells into adipocytes may be manipulated in vitro. 相似文献
105.
The fucose-containing, sulfated polysaccharides from Ascophyllum nodosum and Fucus vesiculosus were isolated by extraction with water adjusted to pH 2. Pure fractions were carefully separated by fractional precipitation with ethanol from aqueous solutions containing magnesium or calcium chloride. Progress in the fractionation efforts and purity of the fractions isolated were established by free-boundary and cellulose acetate clectrophoresis. Ascophyllan, two “complexes”, and a galactofucan were isolated from A. nodosum. An ascophyllan-like fraction, and a “complex” were isolated from F. vesiculosus. Mild, acid hydrolysis (0.02m hydrochloric acid, 1 h, 80°) converted each of the “complexes” into an electrophoretically faster-moving and a slower-moving component. The “complex” from F. vesiculosus comprised a greater proportion of the extract than did the two “complexes” from A. nodosum. In addition, the Fucus “complex” was richer in fucose*. However, the data suggest that neither species contains a pure fucan sulfate in the native state. 相似文献
106.
107.
108.
Mathias Uhlén Björn Nilsson Bengt Guss Martin Lindberg Sten Gatenbeck Lennart Philipson 《Gene》1983,23(3):369-378
Two plasmid vectors, containing the gene coding for staphylococcal protein A and adapted for gene fusion, have been constructed. These vectors will allow fusion of any gene to the protein A gene, thus giving hybrid proteins which can be purified, in a one-step procedure, by IgG affinity chromatography. As an example of the practical use of such vectors, the protein A gene has been fused to the lacZ gene of Escherichia coli. E. coli strains containing such plasmids produce hybrid proteins with both IgG binding and β-galactosidase activities. The hybrid protein(s) can be immobilized on IgG-Sepharose by its protein A moiety with high efficiency without losing its enzymatic activity and they can be eluted from the column by competitive elution with pure protein A. The fused protein(s) also binds to IgG-coated microtiter wells which means that the in vivo product can be used as an enzyme conjugate in ELISA tests. 相似文献
109.
Forslund Kristina Björkman Camilla Abrahamsson Maivor 《Acta veterinaria Scandinavica》1983,24(2):185-199
Plasma cholinesterase (pChE) levels and erythrocyte acetylcholinesterase (eAChE) levels were studied in 6 cows before, during and after parturition (Group I), their calves (Group II), 38 cows suffering from parturient paresis (Group III) and 14 newly delivered non-paretic cows (Group IV). The mean of the pChE level in Group I was 1.5 μkat/1 ± 0.20 before parturition and decreased significantly (P ≦ 0.05) to 1.2 ukat/1 ± 0.16 after parturition. The eAChE level was before parturition ≅ 140 ukat/1 and decreased to ≅ 130 μkat/1 4–5 weeks after parturition. At birth the pChE level was 12.8 ukat/1 ± 5.9 in Group II. After 4 weeks the level had decreased to 2.3 ukat/1 ±0.3. In the bull calves the pChE level started to increase when they were 6 weeks old and reached a level of 5.7 μkat/1 ± 0.6 before slaughter at 6 months of age. The heifers did not show this increase. They had a level of around 2 μkat/1 throughout the investigation. The eAChE level at birth was 119 μkat/1 and increased slowly to a level of 145 μkat/1 at 6 months. No differences between the sexes were found. The cows suffering from parturient paresis had a pChE level of 1.80 μkat/1 ± 0.30 before treatment with calcium (Ca). The level decreased significantly (P ≦ 0.001) after Ca-infusion to a level of 1.67 ukat/1 ±0.29. Group IV had a pChE level of 1.65 μkat/1 ± 0.42 at parturition. Two to 4 months later the cows that had recovered from milk fever had a level of 1.61 μkat/1 ± 0.31 and the control cows 1.66 ukat/1 ± 0.48. No differences between the groups were found for the eAChE level. The findings show that parturition influences the pChE level in cows and that sex influences the pChE level in calves between 6 weeks to at least 6 months of age. Furthermore the elevated pChE level found in the cows suffering from parturient paresis before Ca infusion may be a further sign of a disturbance in the cholinergic system with a special preference to the neuromuscular junctions. 相似文献
110.
Mechanism of intestinal 7 alpha-dehydroxylation of cholic acid: evidence that allo-deoxycholic acid is an inducible side-product 总被引:3,自引:0,他引:3
We previously reported that the 7 alpha-dehydroxylation of cholic acid appears to be carried out by a multi-step pathway in intestinal anaerobic bacteria both in vitro and in vivo. The pathway is hypothesized to involve an initial oxidation of the 3 alpha-hydroxy group and the introduction of a double bond at C4-C5 generating a 3-oxo-4-cholenoic bile acid intermediate. The loss of water generates a 3-oxo-4,6-choldienoic bile acid which is reduced (three steps) yielding deoxycholic acid. We synthesized, in radiolabel, the following putative bile acid intermediates of this pathway 7 alpha,12 alpha-dihydroxy-3-oxo-4-cholenoic acid, 7 alpha,12 alpha-dihydroxy-3-oxo-5 beta-cholanoic acid, 12 alpha-dihydroxy-3-oxo-4,6-choldienoic acid, and 12 alpha-hydroxy-3-oxo-4-cholenoic acid and showed that they could be converted to 3 alpha,12 alpha-dihydroxy-5 beta-cholanoic acid (deoxycholic acid) by whole cells or cell extracts of Eubacterium sp. VPI 12708. During studies of this pathway, we discovered the accumulation of two unidentified bile acid intermediates formed from cholic acid. These bile acids were purified by thin-layer chromatography and identified by gas-liquid chromatography-mass spectrometry as 12 alpha-hydroxy-3-oxo-5 alpha-cholanoic acid and 3 alpha,12 alpha-dihydroxy-5 alpha-cholanoic (allo-deoxycholic acid). Allo-deoxycholic acid was formed only in cell extracts prepared from bacteria induced by cholic acid, suggesting that their formation may be a branch of the cholic acid 7 alpha-dehydroxylation pathway in this bacterium. 相似文献