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During the last few years a growing amount of data has accumulated showing phospholipid par tic i pa tion in nu clear sig nal transduction. Very re cent data strongly sup port the hy poth e sis that sig nal transduction in the nu cleus is au to nomic. Lo cal pro duc tion of inositol polyphosphates, be gin ning with the ac ti va tion of phospholipase C is required for their specific function in the nucleus. Enzymes which modify poly- phosphoinositols may control gene expression. Much less information is available about the role of other lipids in nuclear signal transduction. The aim of this minireview is to stress what is cur rently known about nu clear lipids with re spect to nu clear sig nal transduction.
Trans-4-hydroxy-2-nonenal (4-HNE), a cytotoxic end product of lipid peroxidation, is present in normal human blood plasma at concentrations of 0.1-1.0 μM. It can be, however, further metabolized within a cell, and one of the main products is 4-HNE glutathione conjugate (HNE-SG). In human erythrocyte membrane the system for active extrusion of glutathione (GSH) conjugates of various endo- and xenobiotics has been described; it exhibits either a low (Km at submillimolar concentration range) or a high (Km at low micromolar range) affinity for the transported substrates, such as for example S-(2,4-dinitrophenyl)glutathione (Dnp-SG). In the present study it has been shown that the high affinity transport system for Dnp-SG is competitively inhibited by HNE-SG with Ki of 0.2 μM, while 4-HNE inhibits non-competitively the activity of the transport system for Dnp-SG with Ki of 220 μM. These observations point to the possibility that HNE-SG shares the same transport system with GSH conjugates of other endo- and xenobiotics in erythrocytes. This may be of importance for overall detoxification of the organism under oxidative stress.
We found that in starfish oocytes two different enzymes, phosphatidylserine synthase-1 (PSS1) and -2 (PSS2), which synthesize phosphatidylserine by a base-ex­change reaction, are present. We studied phosphatidylserine synthesis in immature oocytes which still contain the nucleus (germinal vesicles) and in mature cells, in which the re-initiation of the meiotic cycle induced by the hormone 1-methyladenine led to structural changes in the endoplasmic reticulum, to the disappearance of the nu­clear envelope and to the intermixing of the nucleoplasm with the cytoplasm. It was found that the levels of PSS1 and PSS2 transcripts were higher in immature and ma­ture oocytes, respectively. The level of the expressed PSS2 protein, higher than that of PSS1, was not influenced by the maturation process, whereas the level of PSS1 pro­tein was higher in immature than in mature oocytes. Serine incorporation into phosphatidylserine was enhanced in immature oocytes. The depletion of calcium stores by thapsigargin resulted in 50% lowering of phosphatidylserine synthesis. We suggest that changes in phosphatidylserine synthesis may be affected by the release of calcium stored in the nuclear envelope and in the endoplasmic reticulum, the mem­branes that undergo disintegration and fragmentation during meiosis. The reason for the greater synthesis of PS may be the higher level of expression of PSS1 in immature oocytes.
In the present study we investigate the effect of exogenous sphingosine, sphingosine 1-phosphate and sphingosylphosphorylcholine on phospholipase D (PLD) activity in glioma C6 cells. The cells were prelabeled with [1-14C]palmitic acid and PLD-mediated synthesis of [14C]phosphatidylethanol was measured. Sphingosine 1-phosphate and sphingosylphosphorylcholine did not stimulate [14C]phosphatidyl-ethanol formation either at low (0.1-10 μM) or high (25-100 μM) concentrations. On the other hand, sphingosine at concentrations of 100-250 μM strongly stimulated PLD activity as compared to the effect of phorbol ester, 12-O-tetradecanoylphorbol 13-acetate (TPA), known as a PLD activator. The effect of TPA on PLD is linked to the activation of protein kinase C. The present study also shows that sphingosine additively enhances TPA-mediated PLD activity. This is in contrast to the postulated role of sphingosine as a protein kinase C inhibitor. These results demonstrate that in glioma C6 cells sphingosine not only affects PLD independently of its effect on protein kinase C, but also is unable to block TPA-mediated PLD activity.
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