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21.
Control of actin filament length by phosphorylation of fragmin-actin complex 总被引:2,自引:0,他引:2 下载免费PDF全文
Fragmin is a Ca2(+)-sensitive F-actin-severing protein purified from a slime mold, Physarum polycephalum (Hasegawa, T., S. Takahashi, H. Hayashi, and S. Hatano. 1980. Biochemistry. 19:2677-2683). It binds to G-actin to form a 1:1 fragmin/actin complex in the presence of micromolar free Ca2+. The complex nucleates actin polymerization and caps the barbed end of the short F-actin (Sugino, H., and S. Hatano. 1982. Cell Motil. 2:457-470). Subsequent removal of Ca2+, however, hardly dissociates the complex. This complex nucleates actin polymerization and caps the F-actin regardless of Ca2+ concentration. Here we report that this activity of fragmin-actin complex can be abolished by phosphorylation of actin of the complex. When crude extract from Physarum plasmodium was incubated with 5 mM ATP and 1 mM EGTA, the activities of the complex decreased to a great extent. The inactivation of the complex in the crude extract was not observed in the presence of Ca2+. In addition, the activities of the complex inactivated in the crude extract were restored under conditions suitable for phosphatase reactions. We purified factors that inactivated fragmin-actin complex from the crude extract. These factors phosphorylated actin of the complex, and the activities of the complex decreased with an increased level of phosphorylation of the complex. These factors, termed actin kinase, also inactivated the complex that capped the barbed end of short F-actin, leading to elongation of the short F-actin to long F-actin. Thus the length of F-actin can be controlled by phosphorylation of fragmin-actin complex by actin kinase. 相似文献
22.
Actin of fragmin-actin complex is phosphorylated by an endogenous kinase from plasmodium of Physarum polycephalum. The phosphorylation abolishes the nucleation and capping activities of fragmin-actin complex. The kinase has been purified and termed actin kinase [Furuhashi, K. & Hatano, S. (1990) J. Cell Biol. 111, 1081-1087]. Enzymatic properties of the purified actin kinase were studied in detail. Actin kinase exhibited the highest activity under conditions physiological for the plasmodium (30 mM KCl, 6 mM MgCl2, pH 7.0). The Vmax and the Km of the enzyme for ATP were about 83 mumol/min/mg and 25 microM, respectively. The Km for fragmin-actin complex was 190 nM. The purified actin kinase phosphorylated actin of fragmin-actin complex at a constant rate regardless of Ca2+ concentration. Similarly, 2 microM cAMP, 2 microM cGMP, 2 micrograms/ml calmodulin in the presence of Ca2+ or 1 mM GTP showed no effect on the activity of the purified enzyme. Actin kinase did not phosphorylate histone H1, H2B, alpha-casein, or beta-casein, suggesting that actin kinase is a new kind of protein kinase which specifically phosphorylates actin of the fragmin-actin complex. 相似文献
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H. Nakagawa M. Furuhashi T. Umino A. Takago S. Sakamoto 《Zeitschrift fur angewandte Ichthyologie》2004,20(5):389-394
To examine the possibility of dietary α‐starch in reducing feed costs in a practical diet, α‐starch was supplemented at 10, 20, 30 and 40% in a composed diet having the same protein level. The four diets were fed to ayu, Plecoglossus altivelis (initial weight 9.1 g) for 43 days. Growth and feed efficiency increased with the supplement, with values highest in the 30–40%α‐starch diet. The level of dietary α‐starch did not affect the proximate muscle composition; although the hepatosomatic index was not affected, liver glycogen increased with increasing dietary α‐starch. The dietary α‐starch did not influence evacuation time from the gut, and was well digested through passage in the gut, mainly between the stomach and the anterior part of the intestine. Ayu have an ability to adapt their metabolism to high dietary α‐starch, and can digest 40% or more in a composed diet. Although the muscle lipid content did not change, the fatty acid composition was influenced by dietary starch. With the elevation of dietary starch, a decrease of C18:2n‐6 and an increase of C22:6n‐3 occurred. These results indicate that at least 40%α‐starch can be used in practical diets for ayu. 相似文献
25.
A new radioimmunoassay (RIA) for human Chorionic Gonadotropin (hCG) was developed using murine monoclonal antibody to the beta-subunit of hCG (beta-hCG). The IgG fraction of the monoclonal antibody which did not react with 125I-beta-hCG was purified from hybridoma ascites, and covalently coupled to Sepharose 4B. This solid-phase antibody was incubated with standard hCG or serum sampled for 48 hours. The reaction medium was then removed by centrifugation and 125I-beta-hCG and anti-beta-hCG rabbit polyclonal antibody were added to the precipitate. The alcohol precipitation method was used for separating "bound" and "free" forms in the second reaction. The sensitivity for hCG in this assay system was 0.5 mIU/ml serum and the cross-reactivity with human Luteinizing Hormone (hLH) was 0.4%. This assay system was shown to be clinically applicable. Serial serum samples from two patients with trophoblastic disease were assayed and minute amounts of hCG, which could not be determined by conventional assay methods, could be assayed by this new RIA. 相似文献
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Pregnant Wistar rats were injected with bestatin, a specific inhibitor of aminopeptidase M. Placental aminopeptidase M activity was inhibited by injection of bestatin, and fetal body weight was statistically lower than that in the saline-injected or control group. Our present data suggest that placental aminopeptidase M plays an important role in fetal growth. 相似文献
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We investigated the ultrastructural localization of calcium in progenitor predentine and preodontoblasts of developing rat molar tooth germs using the potassium pyroantimonate technique. At the precalcification stage, antimonate reaction product was sparsely, randomly distributed in the preodontoblasts and in the progenitor predentine but no significant reaction could be noticed associated with matrix vesicles. At the matrix vesicle calcification stage, large amounts of antimonate reaction product tended to be localized in the region adjacent to the distal, outer surface membrane of preodontoblasts in which moderate antimonate reaction activity could be observed in mitochondria. Strong antimonate reaction was detected preferentially on the outer surface membrane of some matrix vesicles at this stage. At the subsequent collagen calcification stage, definite antimonate reaction was no longer seen within mitochondria of the late preodontoblasts, instead precipitate was mainly distributed in Golgi area, secretory granules and lateral intercellular spaces. It is suggested that although matrix vesicles contain few calcium capable of reacting to antimonate immediately after their biogenesis, subsequently, large amounts of calcium are accumulated associated with the outer surface membrane of matrix vesicles in the extracellular matrix. 相似文献
28.
Chondroitin sulfate isomers in normal human urine 总被引:1,自引:0,他引:1
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