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21.
The effect of 2,3-diphospho-D-glycerate on the sedimentation coefficient of carbon monoxide hemoglobin was correlated with the fraction of rapidly reacting hemoglobin observed subsequent to flash photolysis at 23 degrees C at pH 7.30 in buffers of 0.1 M ionic strength. Concentrations of the organic phosphate up to about 5 mM resulted in an increase in S20,w, consistent with an increase in the fraction of tetrameric hemoglobin. A decrease in rapidly reacting hemoglobin parallelled the increase in the sedimentation coefficient. Between 5 and 20 mM 2,3-diphosphoglycerate, S20,w decreased, suggesting that dissociation to dimers was enhanced. An increase in rapidly reacting hemoglobin was also observed in this concentration range. Similar sedimentation results were obtained with oxyhemoglobin at pH 7.00 and carbon monoxide hemoglobin at pH 7.06. Assuming single binding sites on each species, the dissociation constants for 2,3-diphosphoglycerate binding to tetrameric and dimeric HbCO are 0.2-0.3 mM and 2-5 mM at pH 7.30. This biphasic effect of this physiologically important organic phosphate on the state of aggregation of R state hemoglobin has not been previously reported, but it is similar to that previously noted with inositol hexaphosphate, which enhanced tetramer formation at low concentrations, while at higher concentrations it promoted hemoglobin dissociation to dimers (White, S. L. (1976) J. Biol. Chem. 251, 4763-4769; Gray, R. D. (1980) J. Biol. Chem. 255, 1812-1818). 相似文献
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Interferon-gamma modulates protein kinase C activity in murine peritoneal macrophages 总被引:21,自引:0,他引:21
T A Hamilton D L Becton S D Somers P W Gray D O Adams 《The Journal of biological chemistry》1985,260(3):1378-1381
The content of Ca2+-, phospholipid-dependent protein kinase activity (protein kinase C) in murine peritoneal macrophages treated with recombinant interferon-gamma (IFN-gamma) has been investigated. Protein kinase C activity was solubilized by nonionic detergent extraction of sonicated cells and separated by high performance liquid chromatography on a TSK 4000 SW gel filtration column. The enzyme eluted from the column in a molecular weight range of 60-80 X 10(3) and was identified by virtue of Ca2+ and phospholipid requirements. Macrophages treated with recombinant IFN-gamma exhibited a substantial increase in total protein kinase activity which could be accounted for entirely by increased protein kinase C activity. This activity was enhanced as much as 5-fold over that seen in untreated macrophages and was specific for IFN-gamma in that other agents known to signal changes in macrophage function had no effect. The time required for the elevation of kinase activity was identical to that required for induction of other functions by IFN-gamma in macrophages. These observations suggest that protein kinase C may be a focus of regulatory action in IFN-gamma-mediated macrophage activation. 相似文献
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Saccharomyces capensis (N.I.U. No. 309) has been shown to have an absolute requirement for inositol and a partial requirement for biotin and pantothenate. Since the relation between growth of this yeast and concentration of inositol is nearly linear in the range of 0.18 to 1.0µg/ml of medium,S. capensis is an excellent organism to use for the bioassay of inositol.
The writers are indebted to Dr. L. R. deMiranda, Centraalbureau voor Schimmelcultures Baarn, the Netherlands for the identification of this yeast. 相似文献
Zusammenfassung Saccharomyces capensis (N.I.U, no. 309) zeigte ein absolutes Erfordernis für Inositol und ein Teilerfordernis für Biotin und Pantothenate. Da die Beziehung zwischen dem Wachstum dieser Hefe und der Konzentration von Inositol beinahe linear im Bereich von 0.18 bis 1.0µg/ml des Nährbodens ist, istS. capensis ein vorzügliches Objekt für Bioassay von Inositol.
The writers are indebted to Dr. L. R. deMiranda, Centraalbureau voor Schimmelcultures Baarn, the Netherlands for the identification of this yeast. 相似文献
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