Site-directed mutagenesis of putative protein kinase C: phosphorylation sites in human AT1 receptor: effects on angiotensin-provoked intracellular calcium mobilisation in stably transfected chinese hamster ovary cells

1997 ◽  
Vol 25 (3) ◽  
pp. 439S-439S ◽  
Author(s):  
STEWART BARKER ◽  
PAUL M. LAVENDER ◽  
ADRIAN J.L. CLARK
Keyword(s):  
Protein Kinase C ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Phosphorylation Sites ◽  
Ovary Cells ◽  
Putative Protein ◽  
Kinase C ◽  
Putative Protein Kinase

scholarly journals Acceleration of Sodium-Calcium Exchange Activity during ATP-induced Calcium Release in Transfected Chinese Hamster Ovary Cells

1997 ◽  
Vol 109 (1) ◽  
pp. 53-60 ◽  
Author(s):  
Melissa Vázquez ◽  
Yu Fang ◽  
John P. Reeves
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Chinese Hamster Ovary ◽  
Calcium Release ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
Kinase C ◽  
Stimulation Of ◽  
Exchange Activity

The P2U purinergic agonist ATP (0.3 mM) elicited an increase in [Ca2+]i due to Ca2+ release from intracellular stores in transfected Chinese hamster ovary cells that express the bovine cardiac Na+/Ca2+ exchanger (CK1.4 cells). The following observations indicate that ATP-evoked Ca2+ release was accompanied by a Ca2+- dependent regulatory activation of Na+/Ca2+ exchange activity: Addition of extracellular Ca2+ (0.7 mM) 0–1 min after ATP evoked a dramatic rise in [Ca2+]i in Na+-free media (Li+ substitution) compared to Na+-containing media; no differences between Na+- and Li+-based media were observed with vector-transfected cells. In the presence of physiological concentrations of extracellular Na+ and Ca2+, the ATP-evoked rise in [Ca2+]i declined more rapidly in CK1.4 cells compared to control cells, but then attained a long-lived plateau of elevated [Ca2+]i which eventually came to exceed the declining [Ca2+]i values in control cells. ATP elicited a transient acceleration of exchange-mediated Ba2+ influx, consistent with regulatory activation of the Na+/Ca2+ exchanger. The acceleration of Ba2+ influx was not observed in vector-transfected control cells, or in CK1.4 cells in the absence of intracellular Na+ or when the Ca2+ content of the intracellular stores had been reduced by prior treatment with ionomycin. The protein kinase C activator phorbol 12-myristate 13-acetate attenuated the exchange-mediated rise in [Ca2+]i under Na+-free conditions, but did not inhibit the ATP-evoked stimulation of Ba2+ influx. The effects of PMA are therefore not due to inhibition of exchange activity, but probably reflect the influence of protein kinase C on other Ca2+ homeostatic mechanisms. We conclude that exchange activity is accelerated during ATP-evoked Ca2+ release from intracellular stores through regulatory activation by increased [Ca2+]i. In the absence of extracellular Ca2+, the stimulation of exchange activity is short-lived and follows the time course of the [Ca2+]i transient; in the presence of extracellular Ca2+, we suggest that the exchanger remains activated for a longer period of time, thereby stabilizing and prolonging the plateau phase of store-dependent Ca2+ entry.

Characterization of Tyrosine-Phosphorylated   Isoform of Protein Kinase C Isolated from Chinese Hamster Ovary Cells

1997 ◽  
Vol 121 (6) ◽  
pp. 1047-1053 ◽  
Author(s):  
M. Kadotani ◽  
T. Nishiuma ◽  
M. Nanahoshi ◽  
Y. Tsujishita ◽  
K. Ogita ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
Kinase C

scholarly journals Concerted action of cytosolic Ca2+ and protein kinase C in receptor-mediated phospholipase D activation in Chinese hamster ovary cells expressing the cholecystokinin-A receptor

1999 ◽  
Vol 337 (2) ◽  
pp. 263-268 ◽  
Author(s):  
Remko R. BOSCH ◽  
Rolf L. L. SMEETS ◽  
Frank SLEUTELS ◽  
Anjana M. P. PATEL ◽  
Sjenet E. Van EMST-De VRIES ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phospholipase D ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
Kinase C ◽  
Peptide Amide ◽  
Sustained Increase

Receptor-mediated activation of phosphatidylcholine phosphatidohydrolase or phospholipase D (PLD) was studied in Chinese hamster ovary (CHO) cells expressing the cholecystokinin-A (CCK-A) receptor. Cells were labelled with [3H]myristic acid for 24 h and PLD-catalysed [3H]phosphatidylethanol formation was measured in the presence of 1% (v/v) ethanol. Cholecystokinin-(26–33)-peptide amide (CCK8) increased PLD activity both time- and dose-dependently. Maximal activation of protein kinase C (PKC) with 1 µM PMA or sustained elevation of the cytosolic free Ca2+ concentration ([Ca2+]i) with 1 µM thapsigargin increased PLD activity to 50% and 70% of the maximal value obtained with CCK8 respectively. The stimulatory effects of CCK8, PMA and thapsigargin were abolished in cells in which PKC was downregulated or inhibited by chelerythrine. PMA/Ca2+-stimulated PLD activity was absent in a homogenate of PKC-downregulated cells but could be restored upon addition of purified rat brain PKC. CCK8-induced PLD activation was inhibited by 90% in the absence of external Ca2+, demonstrating that receptor-mediated activation of PKC in itself does not significantly add to PLD activation but requires a sustained increase in [Ca2+]i. Taken together, the results presented demonstrate that, in CHO-CCK-A cells, receptor-mediated PLD activation is completely dependent on PKC, but that the extent to which PLD becomes activated depends largely, if not entirely, on the magnitude and duration of the agonist-induced increase in [Ca2+]i.

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