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Serine/threonine Protein Phosphatase (threonine + protein_phosphatase)
Selected AbstractsA protein phosphatase 2A from Fagus sylvatica is regulated by GA3 and okadaic acid in seeds and related to the transition from dormancy to germinationPHYSIOLOGIA PLANTARUM, Issue 1 2006Mary Paz González-García Several gibberellic acid (GA3)-induced cDNA fragments encoding putative serine/threonine protein phosphatase (PP) 2A catalytic subunits were obtained by means of differential reverse transcriptase-PCR approach. The full-length clone, named FsPP2A1, isolated from a beechnut cDNA library, exhibited all the features of and homology to members of the PP2A family. By transient expression of FsPP2A1 in tobacco and Arabidopsis cells as a green fluorescent fusion protein, we have obtained evidence supporting the subcellular localization of this protein in both the cytosol and the nucleus. Analysis of FsPP2A1 expression during seed stratification shows that these transcripts increase in the presence of GA3, a treatment proved to be efficient in breaking the dormancy of Fagus sylvatica seeds, but they are almost undetectable in dormant seeds or when dormancy is maintained after treatment with either abscisic acid or the gibberellin biosynthesis inhibitor paclobutrazol. The PP inhibitor okadaic acid (OKA) has a clear effect in decreasing both seed germination and FsPP2A1 expression. Furthermore, FsPP2A1 is specifically expressed in seed tissues, not being detected in other vegetative tissues examined. These results show the regulation of this PP by GA3 and OKA in these seeds. Its relationship with the processes taking place during the transition from dormancy to germination is also discussed. [source] Proteome analysis of human monocytic THP-1 cells primed with oxidized low-density lipoproteinsPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 4 2006Jeong Han Kang Abstract Native low-density lipoprotein (LDL) and oxidized LDL (oxLDL) possess a wide variety of biological properties, and play a central role in atherogenesis. In this study, we used a proteomic analysis of human monocyte THP-1 cells induced with oxLDL or with LDL, to identify proteins potentially involved in atherosclerotic processes. Of the 2500,proteins detected, 93,were differentially expressed as a result of priming with LDL or oxLDL. The proteins were unambiguously identified by comparing the masses of their tryptic peptides with those of all known proteins using MALDI,TOF,MS and the NCBI database. The largest differences in expression were observed for vimentin (94-fold increase), meningioma-expressed antigen,6 (48-fold increase), serine/threonine protein phosphatase,2A (40-fold increase), and beta-1,3-galactosyltransferase (15-fold increase). In contrast, the abundance of an unnamed protein product and phosphogluconate dehydrogenase decreased 30-fold and 25-fold, respectively. The expression of some selected proteins was confirmed by Western blot and RT-PCR analyses. The proteins identified in this study are attractive candidates for further biomarker research. This description of the altered protein profiles induced by oxLDL in human monocytes will support functional studies of the macrophage-derived foam cells involved in the pathogenesis of atherosclerosis. [source] Immunopotentiation on murine spleen lymphocytes induced by polysaccharide fraction of Panax ginseng via upregulating calcineurin activityAPMIS, Issue 4 2010SONG-DONG ZHANG Zhang S-D, Yin Y-X, Wei Q. Immunopotentiation on murine spleen lymphocytes induced by polysaccharide fraction of Panax ginseng via upregulating calcineurin activity. APMIS 2010; 118: 288,96. Calcineurin (CN), a unique Ca2+/calmodulin (CaM)-dependent serine/threonine protein phosphatase, plays a pivotal role in the activation and proliferation of T lymphocytes. Based on the effective molecular screening model established in our laboratory, we found that a part of polysaccharides from the stem and leaves of Panax ginseng, termed PGP-SL, could activate CN activity. Subsequently, we investigated whether PGP-SL also has immunological competence on murine spleen lymphocytes. In the present study, we demonstrated that PGP-SL could significantly promote in vitro spleen lymphocyte proliferation in the absence of either concanavalin A or LPS in a concentration-dependent manner at concentrations ranging from 100 to 500 ,g/ml (p < 0.001). In addition, the proliferation of cyclosporin A (CsA)-treated spleen lymphocytes was also significantly promoted in the same pattern (p < 0.001); the production of IL-2 was elevated and the effect appeared as early as 24 h after PGP-SL treatment. The results of RT-PCR also indicated that the IL-2 mRNA level was markedly enhanced, particularly at PGP-SL concentrations of 300 and 500 ,g/ml, and Fura-2/AM fluorescence probe analysis showed that PGP-SL could dramatically increase the intracellular free calcium concentration of spleen lymphocytes, i.e. [Ca2+]i was significantly increased by approximately 181 and 107% at 300 and 500 ,g/ml of PGP-SL, respectively. However, this effect could be totally inhibited by verapamil treatment. Taking our results together, we suggest that PGP-SL exhibits immunopotentiation effects on murine spleen lymphocytes by the Ca2+,CN,NFAT,IL-2 signaling pathway. [source] Effect of inhibitors of mitogen-activated protein kinase kinase on ,1B -adrenoceptor phosphorylationAUTONOMIC & AUTACOID PHARMACOLOGY, Issue 1-2 2009R. Alcántara-Hernández Summary 1,Mitogen-activated protein kinases mediate hormone/neurotransmitter action on proliferation and differentiation and participate in receptor regulation. The effect of inhibitors of mitogen-activated kinase kinase (MEK) on ,1B -adrenoceptor phosphorylation state and function was studied using different cell lines. It was observed that at nanomolar concentrations the MEK inhibitors, PD98059 (2,-amino-3,-methoxyflavone) and UO126 [1,4-(diamino-2,3-dicyano/1,4-bis-(2-aminophenylthio)-butadiene], increased ,1B -adrenoceptor phosphorylation and diminished the functional response of this receptor to noradrenaline. These agents did not alter the action of lysophosphatidic acid. 2,Staurosporine (IC50 , 0.8 nm) (a general protein kinase inhibitor) and bis-indolyl-maleimide I (IC50 , 200 nm) (a selective protein kinase C inhibitor) inhibited PD98059-induced ,1B -adrenoceptor phosphorylation. In contrast, neither wortmannin (phosphoinositide 3-kinase inhibitor) nor genistein (protein tyrosine kinase inhibitor) had any effect. The data suggest the possibility that MEK might exert control on the activity of the enzymes that regulate receptor phosphorylation, such as G-protein-coupled receptor kinases, protein kinase C or serine/threonine protein phosphatases. 3,Coimmunoprecipitation studies showed a constant association of total extracellular signal-regulated kinase 2 (ERK2) with ,1B -adrenoceptors. Association of phospho-ERK 1/2 to ,1B -adrenoceptors increased not only in response to agonist but also in response to agents that increase ,1B -adrenoceptor and ERK1/2 phosphorylation [such as endothelin-1, phorbol 12-myristate-13-acetate (PMA) and epidermal growth factor (EGF)]; not surprisingly, PD98059 decreased this effect. 4,Our data show that blockade of MEK activity results in increased ,1B -adrenoceptor phosphorylation, diminished adrenoceptor function and perturbation of receptor,ERK1/2 interaction. [source] | ||||||||||