Cellular metabolism regulating H and M currents in bullfrog sympathetic ganglia

1992 ◽  
Vol 70 (S1) ◽  
pp. S51-S55 ◽  
Author(s):  
Takashi Akasu ◽  
Takayuki Tokimasa
Keyword(s):  
Protein Kinase ◽  
Sympathetic Ganglia ◽  
Dependent Manner ◽  
Dependent Protein Kinase ◽  
Maximal Conductance ◽  
Calmodulin Antagonist ◽  
M Current ◽  
Cationic Current ◽  
Voltage Dependent ◽  
Dependent Protein

Much evidence has accumulated suggesting that neurons in autonomic and dorsal root ganglia possess voltage-dependent currents that link with transmitter receptors through intracellular signal transduction systems. The M current (IM), a voltage-dependent potassium current, was activated at potentials more positive than −65 mV, while the H current (IH), a voltage-dependent nonselective cationic current, was activated at potentials more negative than −50 mV. The hydrolyzable form of ATP was required to activate IM and IH. Intracellular application of calmodulin enhanced the amplitude of IM in a calcium-dependent manner. IM was reduced by W-7, a calmodulin antagonist, and by ML-9, an inhibitor of calmodulin-dependent protein kinase. IH was enhanced by intracellular loading with cyclic adenosine monophosphate (AMP) or bath application of forskolin and membrane-permeable cyclic AMP analogues. Isobutylmethylxanthine also increased the maximal conductance of IH. IH was depressed by H-8 but not by phorbol ester. It is concluded that the resting membrane conductance of these ganglion cells can be regulated by basal activities of calmodulin-dependent protein kinase and A kinase.Key words: peripheral neurons, M current, H current, calmodulin, adenylate cyclase.

scholarly journals Ca2+-dependent phosphorylation of tyrosine hydroxylase in PC12 cells.

1985 ◽  
Vol 101 (4) ◽  
pp. 1182-1190 ◽  
Author(s):  
P S Nose ◽  
L C Griffith ◽  
H Schulman
Keyword(s):  
Tyrosine Hydroxylase ◽  
Protein Kinase ◽  
Pc12 Cells ◽  
Cell Function ◽  
Ionophore A23187 ◽  
Dependent Manner ◽  
Dependent Protein Kinase ◽  
Voltage Dependent ◽  
Dependent Protein ◽  
32P Incorporation

Ca2+-dependent protein phosphorylation has been detected in numerous tissues and may mediate some of the effects of hormones and other extracellular stimuli on cell function. In this paper we demonstrate that a Ca2+/calmodulin-dependent protein kinase similar to the enzyme previously purified and characterized from rat brain is present in PC12, a rat pheochromocytoma cell line. We show that Ca2+ influx elicited by various forms of cell stimulation leads to increased 32P incorporation into tyrosine hydroxylase (TH), a major phosphoprotein in these cells. Several other unidentified proteins are either phosphorylated or dephosphorylated as a result of Ca2+ influx. Acetylcholine stimulates TH phosphorylation by activation of nicotinic receptors. K+-induced depolarization stimulates TH phosphorylation in a Ca2+-dependent manner, presumably by opening voltage-dependent Ca2+ channels. Ca2+ influx that results from the direct effects of the ionophore A23187 also leads to TH phosphorylation. Phosphorylation of TH is accompanied by an activation of the enzyme. These Ca2+-dependent effects are independent of cyclic AMP and thus implicate a Ca2+-dependent protein kinase as a mediator of both hormonal and electrical stimulation of PC12 cells.

Up-regulation of functional voltage-dependent sodium channels by cyclic AMP-dependent protein kinase in adrenal medulla

1996 ◽  
Vol 709 (1) ◽  
pp. 37-43 ◽  
Author(s):  
Tomoaki Yuhi ◽  
Akihiko Wada ◽  
Ryuichi Yamamoto ◽  
Toshihiko Yanagita ◽  
Hiromi Niina ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Cyclic Amp ◽  
Sodium Channels ◽  
Adrenal Medulla ◽  
Dependent Protein Kinase ◽  
Voltage Dependent ◽  
Dependent Protein

scholarly journals Ca2+ and phospholipid-dependent protein kinase (protein kinase C) activity is not necessarily required for secretion by human neutrophils

Blood ◽  
1986 ◽  
Vol 68 (4) ◽  
pp. 810-817
Author(s):  
KJ Balazovich ◽  
JE Smolen ◽  
LA Boxer
Keyword(s):  
Protein Kinase ◽  
Phorbol Esters ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Soluble Form ◽  
Human Neutrophils ◽  
Dependent Manner ◽  
Dependent Protein Kinase ◽  
Dependent Protein ◽  
Dose Dependent

Ca2+-dependent and phospholipid-dependent protein kinase (PKC) is a receptor for and is activated by phorbol esters. This enzyme is reportedly involved in the mechanism of superoxide anion (O2-) production and the release of intracellular granule contents from human neutrophils. As previously reported by others, we found that greater than 75% of the total cellular PKC activity existed in a soluble form in untreated neutrophils and that this activity was enhanced in a dose- dependent manner by phorbol 12-myristate 13-acetate (PMA) and by phorbol 12,13-dibutyrate (PDBu). Furthermore, mezerein, an analogue of PMA that is thought to be a competitive inhibitor, did not activate PKC, and on the contrary, inhibited PMA-stimulated activity in a dose- dependent manner. Pretreatment of intact neutrophils with PMA or PDBu caused the “translocation” of PKC activity to the insoluble cell fraction; PKC translocation was not detected after mezerein stimulation at any of the tested concentrations. Neither did mezerein cause an increase in intracellular Ca2+, as monitored by Quin 2 fluorescence. Both phorbol esters and mezerein stimulated intact neutrophils to generate O2- and release lysosomal enzymes into the extracellular medium. Finally sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis demonstrated key differences in the patterns of endogenous phosphoproteins of neutrophils stimulated with phorbol as compared with mezerein. We therefore suggest that PKC activation may not be the only pathway required to elicit neutrophil responses.

iNOS regulation by calcium/calmodulin-dependent protein kinase II in vascular smooth muscle

2007 ◽  
Vol 292 (6) ◽  
pp. H2634-H2642 ◽  
Author(s):  
Rachel J. Jones ◽  
David Jourd'heuil ◽  
John C. Salerno ◽  
Susan M. E. Smith ◽  
Harold A. Singer
Keyword(s):  
Smooth Muscle ◽  
Protein Kinase ◽  
Vascular Smooth Muscle ◽  
Resting Cells ◽  
Dependent Manner ◽  
Dependent Protein Kinase ◽  
Cytokine Induction ◽  
Calcium Calmodulin Dependent ◽  
Protein Kinase Ii ◽  
Dependent Protein

Nitric oxide synthase (NOS) expression is regulated transcriptionally in response to cytokine induction and posttranslationally by palmitoylation and trafficking into perinuclear aggresome-like structures. We investigated the effects of multifunctional calcium/calmodulin-dependent protein kinase II protein kinase (CaMKII) on inducible NOS (iNOS) trafficking in cultured rat aortic vascular smooth muscle cells (VSMCs). Immunofluorescence and confocal microscopy demonstrated colocalization of iNOS and CaMKIIδ2 with a perinuclear distribution and concentration in aggresome-like structures identified by colocalization with γ-tubulin. Furthermore, CaMKIIδ2 coimmunoprecipitated with iNOS in a CaMKII activity-dependent manner. Addition of Ca2+-mobilizing stimuli expected to activate CaMKII; a purinergic agonist (UTP) or calcium ionophore (ionomycin) caused a general redistribution of iNOS from cytosolic to membrane and nuclear fractions. Similarly, adenoviral expression of a constitutively active CaMKIIδ2 mutant altered iNOS localization, shifting iNOS from the cytosolic fraction. Suppression of CaMKIIδ2 using an adenovirus expressing a short hairpin, small interfering RNA increased nuclear iNOS localization in resting cells but inhibited ionomycin-induced translocation of iNOS to the nucleus. Following addition of these chronic and acute CaMKII modulators, there were fewer aggresome-like structures containing iNOS. All of the treatments that chronically affected CaMKII activity or expression significantly inhibited iNOS-specific activity following cytokine induction. The results suggest that CaMKIIδ2 may be an important regulator of iNOS trafficking and activity in VSMCs.

scholarly journals Characterization of ATM Expression, Localization, and Associated DNA-dependent Protein Kinase Activity

1998 ◽  
Vol 9 (9) ◽  
pp. 2361-2374 ◽  
Author(s):  
Dennis P. Gately ◽  
James C. Hittle ◽  
Gordon K. T. Chan ◽  
Tim J. Yen
Keyword(s):  
Dna Damage ◽  
Protein Kinase ◽  
Ataxia Telangiectasia ◽  
Kinase Activity ◽  
Human Fibroblasts ◽  
Protein Kinase Activity ◽  
Dependent Manner ◽  
Dependent Protein Kinase ◽  
Dependent Protein ◽  
Atm Protein

Ataxia telangiectasia–mutated gene (ATM) is a 350-kDa protein whose function is defective in the autosomal recessive disorder ataxia telangiectasia (AT). Affinity-purified polyclonal antibodies were used to characterize ATM. Steady-state levels of ATM protein varied from undetectable in most AT cell lines to highly expressed in HeLa, U2OS, and normal human fibroblasts. Subcellular fractionation showed that ATM is predominantly a nuclear protein associated with the chromatin and nuclear matrix. ATM protein levels remained constant throughout the cell cycle and did not change in response to serum stimulation. Ionizing radiation had no significant effect on either the expression or distribution of ATM. ATM immunoprecipitates from HeLa cells and the human DNA-dependent protein kinase null cell line MO59J, but not from AT cells, phosphorylated the 34-kDa subunit of replication protein A (RPA) complex in a single-stranded and linear double-stranded DNA–dependent manner. Phosphorylation of p34 RPA occurred on threonine and serine residues. Phosphopeptide analysis demonstrates that the ATM-associated protein kinase phosphorylates p34 RPA on similar residues observed in vivo. The DNA-dependent protein kinase activity observed for ATM immunocomplexes, along with the association of ATM with chromatin, suggests that DNA damage can induce ATM or a stably associated protein kinase to phosphorylate proteins in the DNA damage response pathway.

Ca2+/calmodulin-dependent protein kinase II activation and regulation of adrenal glomerulosa Ca2+ signaling

1995 ◽  
Vol 269 (6) ◽  
pp. F751-F760 ◽  
Author(s):  
R. J. Fern ◽  
M. S. Hahm ◽  
H. K. Lu ◽  
L. P. Liu ◽  
F. S. Gorelick ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Kinase Activity ◽  
Low Voltage ◽  
Extracellular Potassium ◽  
Dependent Protein Kinase ◽  
Calmodulin Antagonist ◽  
Cell Extracts ◽  
Protein Kinase Ii ◽  
Dependent Protein ◽  
A Cell

We recently reported that elevations in the intracellular Ca2+ concentration ([Ca2+]i) enhance low-voltage-activated, T-type, Ca2+ channel activity via Ca2+/calmodulin-dependent protein kinase II (CaMKII). Here, we document CaMKII activity in bovine adrenal glomerulosa (AG) cells and assess the importance of CaMKII in depolarization-induced Ca2+ signaling. AG cell extracts exhibited kinase activity toward a CaMKII-selective peptide substrate that was dependent on both Ca2+ [half-maximal concentration for Ca2+ activation (K0.5) = 1.5 microM] and calmodulin (K0.5 = 46 nM) and was sensitive to a calmodulin antagonist and a CaMKII peptide inhibitor. On cell treatment with elevated extracellular potassium (10-60 mM) or angiotensin II, Ca(2+)-independent CaMKII activity increased to 133-205% of basal activity. Ca(2+)-independent kinase activity in agonist-stimulated extracts was inhibited by the CaMKII inhibitor peptide, 1(-)[N,O-bis(1,5- isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (KN-62), a cell-permeable inhibitor of CaMKII, reduced the agonist-induced stimulation of Ca(2+)-independent CaMKII activity. KN-62 also diminished depolarization-induced increases in [Ca2+]i without affecting the membrane potential. These observations suggest that CaMKII is activated in situ by aldosterone secretagogues and augments Ca2+ signaling through voltage-gated Ca2+ channels.

The Role of Calcium/Calmodulin-Dependent Protein Kinase Cascade on MIP-1α Gene Expression in ATL Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3877-3877
Author(s):  
Kensuke Matsumoto ◽  
Koji Murao ◽  
Hitomi Imachi ◽  
Takamasa Nishiuchi ◽  
Hiroaki Ohnishi ◽  
...  
Keyword(s):  
T Cell ◽  
Protein Kinase ◽  
Cell Lines ◽  
Transcriptional Activation ◽  
Promoter Activity ◽  
Calcium Nitrate ◽  
Dependent Manner ◽  
Dependent Protein Kinase ◽  
Dependent Protein

Abstract Adult T-cell leukemia (ATL) is a mature CD4+ T-cell malignancy caused by infection with human T-lymphotrophic virus (HTLV-1) and is associated with a marked hypercalcemia on many patients. Recently, it is proposed that Macrophage inflammatory protein-1α (MIP-1α) is the clinical hallmark of hypercalcemia in ATL, but the regulation of MIP-1α secretion has not been clarified yet. In this study, we examined the effect of calcium on the MIP-1α secretion and cell proliferation of ATL cells, and also the role of the Ca2+/calmodulin (CaM)-dependent protein kinase (CaM-K) cascade in transcriptional activation of MIP-1α. The addition of calcium nitrate to the medium enhanced the secretion of MIP-1α by ATL cell lines (ATN, MT-2, OKM-2T) and the proliferation in a dose dependent manner. The maximum response of MIP-1α secretion was induced at 10.42mM calcium in OKM-2T cells. CaM-KK selective inhibitor STO-609 inhibited the calcium dependent secretion of MIP-1α and proliferation of ATL cell lines. We investigated the effects of CaM-KK/CaM-KIV signaling pathway on MIP-1α promoter activity in OKM-2T. The transfection of CaM-KIV stimulated the MIP-1α promoter activity and the upstream kinase, CaM-KK enhanced the stimulatory effect of CaM-KIV on its activity. Furthermore, mutation of the cAMP response element (CRE) within the MIP-1α promoter significantly reduced the effect of CaM-KIV and calcium, and it wasn’t less enhanced by the addition of calcium nitrate to the medium than the wild type. Our studies have indicated that hypercalcemia enhances MIP-1α secretion and the cell growth in ATL cells, and these mechanisms require the CaM-KK/CaM-KIV cascade. These findings raise the possibility that the inhibitory of CaM-KK/CaM-KIV cascade may be of therapeutic value for ATL.

scholarly journals Regulation of prostaglandin synthesis by protein kinase C in mouse peritoneal macrophages

1989 ◽  
Vol 260 (2) ◽  
pp. 471-478 ◽  
Author(s):  
H J Pfannkuche ◽  
V Kaever ◽  
D Gemsa ◽  
K Resch
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Peritoneal Macrophages ◽  
Dependent Manner ◽  
Membrane Phospholipids ◽  
Dependent Protein Kinase ◽  
Kinase C ◽  
Dependent Protein ◽  
Dose Dependent ◽  
Mouse Peritoneal Macrophages

Resident mouse peritoneal macrophages synthesized and released prostaglandins (PGs) when challenged with 12-O-tetradecanoylphorbol 13-acetate (TPA) or 1,2-dioctanoyl-sn-glycerol (DiC8). Both stimuli were found to activate Ca2+/phospholipid-dependent protein kinase C (PKC). 1-(5-Isoquinolinesulphonyl)-2-methylpiperazine (‘H-7’) and D-sphingosine, known to inhibit PKC by different mechanisms, were able to decrease the PKC activity of macrophages in a dose-dependent manner. Addition of either PKC inhibitor decreased PG synthesis and also the release of arachidonic acid (AA) from phospholipids induced by TPA or DiC8. Simultaneously TPA or DiC8 also decreased incorporation of free AA into membrane phospholipids of macrophages. AA incorporation could be restored, however, by pretreatment with the PKC inhibitors. Our results demonstrate an involvement of PKC in the regulation of PG synthesis in mouse peritoneal macrophages and provide further evidence that reacylation of released fatty acids may be an important regulatory step.

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