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121.
In extracts from Zea mays shoots, the presence of thiol compoundsin the extraction buffer was necessary to get an active 3 deoxy-D-arabinoheptulosonic acid 7-phosphate (DAHP) synthase. Its pH optimumfor activity was about 7.5. Of the different cations tested,only Mn++ was an activator. Enzyme stability was optimal inTris-HCl buffer, pH 7.5, that contained a reducing agent, Mn++and a polyol. Contrary to other reports, phosphoenolpyruvate(PEP) did not stabilize the preparation significantly. The synthaseexhibited high affinities for both erythrose-4-phosphate (Km:0.24 mM) and PEP (Km: 0.31 mM). Its specific activity was highestin young shoots. Corn DAHP synthase was inhibited in vitro by tryptophan. Moreover,the enzyme was retarded on a tryptophan agarose affinity column,but it was removed with the bulk of protein from the same supportwhen eluted with buffer containing tryptophan. Inhibition whichwas easily lost during storage at 4°C was pH dependent andincreased during development. Maximal inhibition, about 60%with 1 mM tryptophan, was observed in extracts from 8 day-oldshoots. Phenylalanine and tyrosine were not inhibitory, andno synergistic effects were observed when the aromatic aminoacids were tested in combination. Isoenzymes could not be demonstrated. (Received April 23, 1980; ) 相似文献
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Alain B. Schreiber Johan Hoebeke Bernard Vray A. Donny Strosberg 《Experimental cell research》1981,132(2)
The association and dissociation mechanisms of lectin membrane receptor microclustering on HeLa cells have been studied by measuring resonance energy transfer between fluoresceinated and rhodaminated lentil lectin. Compounds known to affect membrane receptor mobility, such as Ca2+ ions, methylamine, cytochalasin D and nocodazole, did not modify the association kinetics nor the maximal energy transfer values at 4 and 37 °C. Dissociation of the membrane receptor microclusters was followed by measuring the temporal decrease in energy transfer values at 4 °C after preincubation for different time intervals at 37 °C. The rate of dissociation of the lectin receptors decreased in the presence of Ca2+ ions (10−3 M) and after cross-linking with anti-lectin antibodies. An increase was observed in the presence of cytochalasin D (10−6 M) and, to a lesser extent, of methylamine (10−2 M). When cytochalasin D and methylamine were combined at subliminal concentrations, a partial synergistic effect was observed. Nocodazole (10−6 M) had no effect. The results suggest that the association of lectin membrane receptors in microclusters is mediated only by physicochemical parameters. Ca2+ ions, cytochalasin D (microfilaments) and methylamine (transglutaminase)-sensitive components appear, however, to play an important role in the stabilization of the receptor microclusters. 相似文献
124.
Using mouse thymocytes, mitogen-induced [3H]thymidine incorporation was compared with a recently developed flow-cytometric technique, based on acridine orange staining of cells, which differentiates the G0 and G1 phase of thymocytes. PHA induces a transient but considerable G0-G1 shift without any substantial proliferation. On the other hand, crude supernatants derived from Con A-stimulated human peripheral blood mononuclear cells induce only a minor G0-G1 shift and no proliferation. However, PHA in the presence of this supernatant induced an increased [3H]thymidine uptake in thymocytes and a shift from G1 to S. These results support the current hypothesis that a factor present in Con A-activated supernatants in conjunction with PHA stimulation indeed facilitates the entrance of G1 cells into the S phase. The flow-cytometric technique might be used in the study of the interaction of endogenous mediators with exogenous mitogenic agents in activating lymphocytes to proceed through the initial G0-G1 phases of the cell cycle. 相似文献
125.
Four strains of both Taphrina pruni and T. institiae were cultivated under identical conditions and and lipids and fatty acids were quantitatively analysed at two stages of their development. Tri- and diglycerides are the major neutral lipids in both species. Phosphatidylcholine and phosphatidylethanolamine are the most abundant polar lipids. Qualitatively, the two species show identical fatty acid contents, except for margaric acid (17:0) which was only found in Taphrine pruni. Quantitatively there are several differences: palmitoleic acid (16:1) occurs in reasonable amounts regularly and only in Taphrina pruni. The ratios 16:0/18:0 and 18:1/18:2 are generally higher for T. insititiae whereas the degree of unsaturation of fatty acids is higher in the former. The results are discussed with regard to data on other fungal species. 相似文献
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Abstract Plasmid pIP501 was transferred by conjugation from Lactococcus lactis to Lactobacillus delbrückii subsp. bulgaricus and Lactobacillus helveticus . Only Lb. delbrückii subsp. bulgaricus transconjugants could act as a donor in crosses with Lc. lactis . No Lactobacillus transconjugants were detected after inter- or intra-species Lactobacillus crosses. Plasmid pIP501 has undergone no detectable deletion or rearrangement during transfer from Lc. lactis to Lactobacillus strains. 相似文献
130.