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Conventional PKC (conventional + pkc)

Distribution by Scientific Domains

Life Sciences	50%
Medical Sciences	33%

Selected Abstracts

Bimodal role of conventional protein kinase C in insulin secretion from rat pancreatic , cells

THE JOURNAL OF PHYSIOLOGY, Issue 1 2004
Hui Zhang
The present study was conducted to evaluate the role of conventional protein kinase C (PKC) in calcium-evoked insulin secretion. In rat , cells transfected with green fluorescent protein-tagged PKC-, (PKC-,,EGFP), a depolarizing concentration of potassium induced transient elevation of cytoplasmic free calcium ([Ca2+]c), which was accompanied by transient translocation of PKC-,,EGFP from the cytosol to the plasma membrane. Potassium also induced transient translocation of PKC-,,EGFP, the C1 domain of PKC-, and PKC-,,GFP. A high concentration of glucose induced repetitive elevation of [Ca2+]c and repetitive translocation of PKC-,,EGFP. Diazoxide completely blocked both elevation of [Ca2+]c and translocation of PKC-,,EGFP. We then studied the role of conventional PKC in calcium-evoked insulin secretion using rat islets. When islets were incubated for 10 min with high potassium, Gö-6976, an inhibitor of conventional PKC, and PKC-, pseudosubstrate fused to antennapedia peptide (Antp-PKC19,31) increased potassium induced secretion. Similarly, insulin release induced by high glucose for 10 min was enhanced by Gö-6976 and Antp-PKC19,31. However, when islets were stimulated for 60 min with high glucose, both Gö-6976 and Antp-PKC19,31 reduced glucose-induced insulin secretion. Similar results were obtained by transfection of dominant-negative PKC-, using adenovirus vector. Taken together, PKC-, is activated when cells are depolarized by a high concentration of potassium or glucose. Conventional PKC is inhibitory on depolarization-induced insulin secretion per se, but it also augments glucose-induced secretion. [source]

Enhancement of anchorage-independent growth of human pancreatic carcinoma MIA PaCa-2 cells by signaling from protein kinase C to mitogen-activated protein kinase

MOLECULAR CARCINOGENESIS, Issue 4 2002
Keiko Ishino
Abstract We found that 12- O -tetradecanoylphorbol-13-acetate (TPA) promoted anchorage-independent growth but did not affect anchorage-dependent growth of MIA PaCa-2 human pancreatic carcinoma cells. TPA markedly activated mitogen-activated protein kinase (MAPK)/extracellular signal,regulated kinase in an anchorage-independent manner. Two protein kinase C (PKC) isoforms, conventional PKC (cPKC) and novel PKC (nPKC), but not apical PKC, translocated from the cytosolic to the particulate fraction upon TPA treatment. To identify the PKC isoforms involved in the regulation of anchorage-independent growth, four PKC isoforms (,, ,, ,, and ,) were forced to be expressed in MIA PaCa-2 cells with an adenovirus vector. Overexpression of nPKC, or nPKC, activated MAPK and promoted anchorage-independent growth. Overexpression of cPKC, alone did not influence anchorage-independent growth but lowered the concentration of TPA that was required to enhance such growth. Expression of constitutively active MAPK kinase-1 (MEK1) also promoted anchorage-independent growth. Furthermore, PKC inhibitors or an MEK inhibitor completely suppressed both TPA-induced activation of MAPK and promotion of anchorage-independent growth, but a cPKC-selective inhibitor partially suppressed TPA-induced promotion of the growth. Based on these results, we suggest that MAPK activation, mediated by certain isoforms of PKC, plays a part in oncogenic growth of MIA PaCa-2 cells. In summary, our data indicated that specific inhibitors of the cPKC and nPKC signaling pathway might be selective anti-oncogenic growth agents for some types of human pancreatic cancer. © 2002 Wiley-Liss, Inc. [source]

Bimodal role of conventional protein kinase C in insulin secretion from rat pancreatic , cells

Possible role of the protein kinase C/CPI-17 pathway in the augmented contraction of human myometrium after gestation

BRITISH JOURNAL OF PHARMACOLOGY, Issue 7 2003
Hiroshi Ozaki
Activation of protein kinase C (PKC) by phorbol 12,13-dibutylate (PDBu, 1 ,M) induced sustained contractions with no increase in [Ca2+]i in nonpregnant and pregnant human myometria. The contractile effects of PDBu in pregnant myometrium were much greater than those in nonpregnant myometrium, and the contractions in pregnant myometrium were accompanied by an increase in myosin light chain (MLC) phosphorylation at Ser19. The contraction induced by PDBu in pregnant myometrium was inhibited by the inhibitors of conventional PKC isoforms, bisindolylmaleimides and indolocarbazole, such as Go6976, Go6983, and Go6850 (1 ,M). LY333531 (1 ,M), a specific inhibitor of PKC,, also inhibited the PDBu-induced contraction in the pregnant myometrium. In the pregnant myometrium permeabilized with , -toxin, PDBu increased the contractions induced at fixed Ca2+ concentration (0.3 ,M) both in nonpregnant and pregnant myometria, indicating Ca2+ sensitization of contractile elements. Western immunoblot analysis indicated that pregnant myometrium contained PKC isozymes such as conventional PKC (,, ,, ,), novel PKC (,, ,, ,), and atypical PKC (, but not , and ,). RT-PCR and real-time RT-PCR analysis indicated that, among the conventional PKC, the levels of mRNA of , isoform in pregnant human myometrium were greater than those in nonpregnant myometrium. CPI-17 is a substrate for PKC, and the phosphorylated CPI-17 is considered to inhibit myosin phosphatase. The levels of CPI-17 mRNA and protein expression were also greater in the pregnant myometrium. These results suggest that the PKC-mediated contractile mechanism is augmented in human myometrium after gestation, and that this augmentation may be attributable to the increased activity of the , PKC isoform and CPI-17. British Journal of Pharmacology (2003) 140, 1303,1312. doi:10.1038/sj.bjp.0705552 [source]

Role of atypical protein kinase C isozymes and NF-,B in IL-1,-induced expression of cyclooxygenase-2 in human myometrial smooth muscle cells

JOURNAL OF CELLULAR PHYSIOLOGY, Issue 3 2007
Sara V. Duggan
Increased myometrial expression of cyclooxygenase-2 (Cox-2) at term results from elevated local levels of inflammatory cytokines, and its inhibition provides a potential route for intervention in human pre-term labor. We have identified a role for atypical protein kinase C (PKC) isozymes in IL-1,-induced Cox-2 expression in human myometrial smooth muscle cells (HMSMC). The PKC inhibitor GF109203X (10 µM) inhibited IL-1,-induced Cox-2 protein and RNA expression, which were also reduced by MAPK and nuclear factor ,B (NF-,B) inhibitors. GF109203X did not affect MAPK activities, and neither did it replicate the effect of p38 MAPK inhibition on Cox-2 mRNA stability, suggesting that PKC operates through an independent mechanism. The effect of GF109203X remained intact after depletion of conventional and novel PKC isozymes by phorbol ester pre-treatment. In contrast LY379196 (10 µM), which at micromolar concentrations inhibits all but atypical PKCs, did not affect Cox-2 expression. A peptide corresponding to the pseudosubstrate sequence of atypical PKCs blocked Cox-2 protein expression, whereas the sequence from conventional PKCs was ineffective. GF109203X did not affect NF-,B binding to nuclear proteins, but strongly reduced NF-,B-dependent transcription in luciferase reporter assays. Our findings indicate that IL-1,-induced Cox-2 expression in HMSMC in culture requires p38-MAPK-mediated mRNA stabilization and an independent activation of Cox-2 transcription which is dependent on the action of atypical PKCs, probably through direct stimulation of the transactivating activity of NF-,B. J. Cell. Physiol. 210: 637,643, 2007. © 2006 Wiley-Liss, Inc. [source]

Quantitative proteome analysis of detergent-resistant membranes identifies the differential regulation of protein kinase C isoforms in apoptotic T cells

PROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 15 2010
Therese Solstad
Abstract Several lines of evidence suggest that detergent-resistant membranes (DRMs) (also known as lipid rafts and glycosphingolipid-enriched microdomains) may have a role in signaling pathways of apoptosis. Here, we developed a method that combines DRMs isolation and methanol/chloroform extraction with stable isotope labeling with amino acids in cell culture-based quantitative proteome analysis of DRMs from control and cisplatin-induced apoptotic Jurkat T cells. This approach enabled us to enrich proteins with a pivotal role in cell signaling of which several were found with increased or decreased amounts in DRMs upon induction of apoptosis. Specifically, we show that three isoforms of protein kinase C (PKC) are regulated differently upon apoptosis. Although PKC, which belongs to the group of conventional PKCs is highly up-regulated in DRMs, the levels of two novel PKCs, PKC, and PKC,, are significantly reduced. These alterations/differences in PKC regulation are verified by immunoblotting and confocal microscopy. In addition, a specific enrichment of PKC, in apoptotic blebs and buds is shown. Furthermore, we observe an increased expression of ecto-PKC, as a result of exposure to cisplatin using flow cytometry. Our results demonstrate that in-depth proteomic analysis of DRMs provides a tool to study differential localization and regulation of signaling molecules important in health and disease. [source]