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1

Exogenous sphingosine 1-phosphate and sphingosylphosphorylcholine do not stimulate phospholipase D in C6 glioma cells

100%
Dygas A., Sidorko M., Bobeszko M., Baranska J.
Acta Biochimica Polonica
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1999
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tom 46
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nr 1
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.
2

The transition from a pharmacophore-guided approach to a receptor-guided approach in the design of potent protein kinase C ligands

86%
Marquez V. E., Nacro K., Benzaria S., Lee J., Milne G. W., Bienfait B., Wang S., Lewin N., Blumberg P. M.
Cellular and Molecular Biology Letters
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1998
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tom 03
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nr 3
3

Regulation of mammalian phospholipase D by ARF, RHOA, and an unidentified protein factor

86%
Nakamura S.
Cellular and Molecular Biology Letters
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1997
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tom 02
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nr 2
4

Fluorometric assay of oleate-activated phospholipase D isoenzyme in membranes of rat nervous tissue and human platelets

72%
Krzystanek M., Trzeciak H. I., Krzystanek E., Malecki A.
Acta Biochimica Polonica
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2010
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tom 57
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nr 3
 Phospholipase D plays a key role in the biosynthesis of phosphatidic acid, a second messenger involved in essential cellular processes. Oleate-activated phospholipase D was the first mammalian phospholipase D isoform to be discovered but is the least known. The study was aimed to test a fluorometric method of assessment of oleate-activated phospholipase D activity in different biological materials. The brain cortex of male Wistar rats, cultured rat brain astrocytes, and human platelets were processed to yield plasmatic membranes for experiments. To assess phospholipase D activity the modified fluorometric method was used. Previously, the method was used only to determine H2O2. In this enzyme-coupled assay phospholipase D activity is monitored indirectly using 10-acetyl-3,7-dihydroxyphenoxazine. First, phospholipase D cleaves exogenous phosphatidylcholine to yield choline and phosphatidic acid. Second, choline is oxidized by choline oxidase to betaine and H2O2. Finally, in the presence of horseradish peroxidase, H2O2 reacts with 10-acetyl-3,7-dihydroxyphenoxazine to generate the highly fluorescent product, resorufin. The concentration of resorufin was measured using excitation and emission at 560 nm and 590 nm, respectively. The proposed optimal parameters of the tested assay are 25 μg of rat brain cortex protein, 50 μg of rat brain astrocyte protein, and 50 μg of human platelet protein in a reaction volume of 200 μL, and 2 min enzymatic reaction at 37°C. The fluorometric method may be applied to assay phospholipase D in different biological materials.
5
Content available remote

Screening and modulation of extracellular signals by mucous barrier. Serum glycosylphosphatidylinositol phospholipase D (GPI-PLD) releases protective mucous barrier from oral mucosa

72%
Slomiany A., Nishikawa H., Slomiany B. L.
Journal of Physiology and Pharmacology
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2002
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tom 53
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nr 1
Performance of mucosal epithelial barrier is modified by numerous agents that exert effects on mucin-Mucin Binding Protein (MBP)complex.The aim of the studies described was to determine the nature of the damage or modification of oral mucous barrier by the short-term exposure to ethanol.Methods:Culture of rat buccal mucosa in the presence of ethanol and [3H]-labeled proline and palmitate revealed substantial decrease in MBP synthesis and the release of MBP to the medium.The radioscanning of the samples prepared from the culture medium and the apical epithelial membranes subjected to SDS- PAGE and western blotting disclosed that the released,water soluble 97kDa MBP glycopeptide was labeled with proline and palmitate.When the experiments were conducted in the presence of 5mM EDTA,the GPI-PLD inhibitor,the majority of radiolabeled MBP remained in the membrane-bound form and was extractable with Triton X-114.The results on the purified GPI-linked MBP degradation by serum enzyme,by the saliva containing serum transudate,and the suppression of the process by inclusion of GPI-PLD-specific inhibitor support our contention that membrane MBP is released to medium by GPI-PLD- like activity.Results:The release of MBP from apical epithelial surfaces was induced by depletion of mucin and the presence of serum-derived GPI-PLD in the tissue homogenate. Strong likelihood exists that under in situ conditions ethanol-induced transudation of serum to saliva provides the vehicle for the transfer of GPI-PLD activity to salivary contents. Defacement of the oral surfaces from mucous barrier signals prospect of lumenal agent influence on the unprotected epithelial exterior,and allows ingression of microbes and untoward acting substances into the organism.
6

A new BSMV-based vector with modified beta molecule allows simultaneous and stable silencing of two genes

58%
Kawalek A., Dmochowska-Boguta M., Nadolska-Orczyk A., Orczyk W.
Cellular and Molecular Biology Letters
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2012
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tom 17
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nr 1
Virus-induced gene silencing is an important tool for functional gene analysis and the vector based on Barley stripe mosaic virus (BSMV) is widely used for the purpose in monocots. Of the tripartite BSMV genome, currently the BSMV:γMCS molecule is used to clone a fragment of a target gene. As an alternative, the BSMV:β molecule was engineered with a unique BamHI site between the open reading frame of βc (ORF βc) and poly(A). The mixture of RNA particles α, βBamHI and γMCS was fully infectious. Barley phytoene desaturase and wheat phospholipase Dα fragments were cloned to βBamHI and γMCS. Delivery of the target gene fragment in γMCS induced stronger silencing, while delivery in βBamHI yielded more stable transcript reduction. A quantitative analysis (qRT-PCR) of the transcripts showed that the silencing induced with a fragment carried in both particles was stronger and more stable than that from a fragment placed in one particle. The modification of β enables simultaneous silencing of two genes. Quantifying the β and γ particles in virus-inoculated plants revealed a 2.5-fold higher level of γ than β, while the stability of the insert was higher in β compared with γ. The possible influence of the relative quantity of β and γ particles in virus-inoculated plants on insert stability and gene silencing efficiency is discussed.
7
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Content available

Phospholipase D mediated transphospatidylation as a possible new pathway of ethanol metabolism in isolated rat pancreatic acini

58%
Rydzewska G., Jurkowska G., Gabryelewicz A.
Journal of Physiology and Pharmacology
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1996
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tom 47
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nr 2
8
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Content available

The influence of polyamines synthesis inhibition on pancreas regeneration and phospholipase D activity after acute caerulein induced pancreatitis in rats. Biochemical and ultrastructural study

58%
Jurkowska G., Rydzewska G., Andrzejewska A.
Journal of Physiology and Pharmacology
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1997
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tom 48
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nr 4
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