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101.
A membrane component involved in the transport of adenosine in adipocytes has been identified utilizing the techniques of photoaffinity labeling with the adenosine derivative, 8-azidoadenosine. In the absence of light, adenosine and 8-azidoadenosine exhibited similar transport characteristics. In addition, adenosine was shown to be a competitive inhibitor of 8-azidoadenosine uptake, and the photoprobe, a competitive inhibitor of adenosine uptake. Analysis of the nucleotide metabolites indicated that the photoprobe was metabolized in a similar fashion to that observed for adenosine. Several nucleoside transport inhibitors were also equally effective in inhibiting the uptake of both nucleosides. These results suggest that 8-azidoadenosine is transported by the same membrane system as adenosine. Photolysis of 8-azido[2-3H]adenosine in the presence of adipocytes resulted in the covalent incorporation of the photoprobe into the plasma membrane fraction. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that essentially all of the radioactivity was incorporated into a glycoprotein with a molecular weight of 56,000. This labeling was inhibited by greater than 90% when the photolysis was carried out in the presence of excess adenosine or the transport inhibitors, persantin or theophylline. Fractionation of the labeled plasma membranes by dialysis against water (pH 9.5) indicated that approximately 75% of the radioactivity was associated with a glycoprotein which resisted solubilization by this procedure. These results suggest that the major labeled species is a 56,000 Mr intrinsic membrane glycoprotein which may function as a component of a transmembrane assembly involved in the transport of adenosine. 相似文献
102.
103.
E Pineda-Molina P Klatt J Vázquez A Marina M García de Lacoba D Pérez-Sala S Lamas 《Biochemistry》2001,40(47):14134-14142
The cellular redox status can modify the function of NF-kappaB, whose DNA-binding activity can be inhibited by oxidative, nitrosative, and nonphysiological agents such as diamide, iodoacetamide, or N-ethylmaleimide. This inhibitory effect has been proposed to be mediated by the oxidation of a conserved cysteine in its DNA-binding domain (Cys62) through unknown biochemical mechanisms. The aim of this work was to identify new oxidative modifications in Cys62 involved in the redox regulation of the NF-kappaB subunit p50. To address this problem, we exposed p50, both the native form (p50WT) and its corresponding mutant in Cys62 (C62S), to changes in the redox pair glutathione/glutathione disulfide (GSH/GSSG) ratio ranging from 100 to 0.1, which may correspond to intracellular (patho)physiological states. A ratio between 1 and 0.1 resulted in a 40-70% inhibition of the DNA binding of p50WT, having no effect on the C62S mutant. Mass spectrometry studies, molecular modeling, and incorporation of (3)H-glutathione assays were consistent with an S-glutathionylation of p50WT in Cys62. Maximal incorporation of (3)H-glutathione to the p50WT and C62S was of 0.4 and 0.1 mol of (3)H-GSH/mol of protein, respectively. Because this covalent glutathione incorporation did not show a perfect correlation with the observed inhibition in the DNA-binding activity of p50WT, we searched for other modifications contributing to the maximal inhibition. MALDI-TOF and nanospray-QIT studies revealed the formation of sulfenic acid as an alternative or concomitant oxidative modification of p50. In summary, these data are consistent with new oxidative modifications in p50 that could be involved in redox regulatory mechanisms for NF-kappaB. These postranslational modifications could represent a molecular basis for the coupling of pro-oxidative stimuli to gene expression. 相似文献
104.
105.
106.
Jean M. Karle Lawrence W. Anderson Daniel D. Dietrick Richard L. Cysyk 《Analytical biochemistry》1980,109(1):41-46
Serum and plasma uridine levels in mice, rats, and humans were conveniently measured by reverse-phase high-pressure liquid chromatography. Human serum levels were in the range of 1.9 to 8.4 nmol/ml, rat serum levels ranged from 3.7 to 9.4 nmol/ml, and mouse serum levels measured 8.0 to 11.8 nmol/ml. Levels of uridine were the same in human plasma and serum; however, plasma from mice and rats was somewhat lower than serum in uridine content ranging from 1.7 to 4.1 nmol/ml in rats and 1.5 to 4.7 nmol/ml in mice. There was some variation in the individual human serum and rat plasma uridine levels throughout the day, but the values were within the normal range, and the variations had no set pattern. Withholding food for 16–24 h had no observable effect on serum uridine levels in mice and humans or on rat plasma levels. These results suggest that uridine levels are regulated and are not a direct reflection of dietary intake of uridine. 相似文献
107.
Differences in the rabbit uterine response to progesterone as influenced by growth hormone or prolactin 总被引:1,自引:0,他引:1
The direct effect of growth hormone (GH) on the uterine response to progesterone was tested by using ovariectomized rabbits (at least 12 weeks) treated with GH; GH + progesterone; or progesterone alone. These results were compared with the effect of prolactin or prolactin + progesterone on the uterus. Prolactin treatment produced an increase (P less than 0.01) in the endometrial surface area and restored cytosolic oestrogen and progesterone receptor concentrations to oestrous control values. The sequential treatment of does with prolactin + progesterone stimulated uteroglobin production to a concentration equal to that found in intact rabbits on Day 5 of pregnancy. In contrast, GH treatment had no effect on endometrial surface area, produced an increase in the concentration of cytosolic oestrogen receptor but did not produce an increase in the concentration of progesterone receptor. The sequential treatment of does with GH + progesterone failed to stimulate uteroglobin secretion above control (progesterone alone) values. It is concluded that the action of prolactin in the rabbit uterus is no generally somatogenic; rather, prolactin increases the concentration of progesterone receptor and thereby enhances the uterine response to progesterone. 相似文献
108.
F García J Sánchez J Planas 《Comparative biochemistry and physiology. A, Comparative physiology》1987,87(4):947-950
1. Male quails submitted 20 and 120 days to a low iron diet (7 ppm) were compared to female laying quails, exposed for 30 days to the same low iron regime, in order to compare the response of the iron metabolic control under a single (erythropoiesis) or a doubled (erythropoiesis and egg formation) iron demand. 2. Iron deposit in storage organs, the classical hematology and the intestinal iron absorption were analyzed in these animals. 3. In males, after 120 days, the iron deposits were reduced 50 and 75%, but hematological values (hematocrit and hemoglobin concentration) were normal, although in laying quails, after 30 days, an anemic condition was evident in both blood parameters and iron deposits, provoking an iron deficient erythropoiesis. 4. The enhancement of the intestinal iron uptake, confirms the anemic character of these birds. 相似文献
109.
110.
Daniel A. Vardy Csaba Kari Gerald S. Lazarus Pamela J. Jensen Asher Zilberstein Gregory D. Plowman Ulrich Rodeck 《Journal of cellular physiology》1995,163(2):257-265
Autocrine activation of the epidermal growth factor (EGF) receptor on keratinocytes has been recognized as an important growth regulatory mechanism involved in epithelial homeostasis, and, possibly, hyperproliferative diseases. Insulin-like growth factor (IGF)-1 and insulin have been shown to be paracrine keratinocyte mitogens that bind to the type I IGF receptor which is expressed on actively proliferating keratinocytes in situ. In this report, we demonstrate that IGF-1/insulin induced production of keratinocyte-derived autocrine growth factors that bind to the EGF receptor. Increased steady-state mRNA levels for transforming growth factor alpha (TGF-α) and for amphiregulin (AR) were observed upon incubation of keratinocytes with mitogenic concentrations of IGF-1. IGF-1 also induced production and secretion of TGF-α and AR proteins as detected by immunoassays. An EGF receptor antagonistic monoclonal antibody abolished the mitogenic effect of IGF-1 on cultured keratinocytes. These results suggest that stimulation of keratinocyte growth by IGF-1 requires activation of an EGF receptor-mediated autocrine loop. © 1995 Wiley-Liss, Inc. 相似文献