Calcium and protein kinase C (PKC)-related kinase mediate α1A-adrenergic receptor-stimulated activation of phospholipase D in rat-1 cells, independent of PKC

Jean Hugues Parmentier, Aftab Ahmed, Ying Ruan, Gautam K. Gandhi, Abdelwahab E. Saeed, Kafait Malik

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

A previous study conducted in rat-1 cells expressing α1A-adrenergic receptors showed that phenylephrine (PHE) stimulates phospholipase D (PLD) activity. This study was conducted to determine the contribution of protein kinase C (PKC) to PHE-induced PLD activation in these cells. PKC inhibitors bisindolylmaleimide (BIM) I and Ro 31-8220, but not Gö 6976 or a pseudosubstrate peptide inhibitor of PKCα, decreased PLD activity and arachidonic acid release elicited by PHE. However, antisense oligonucleotides directed against PKC α, δ, ε, and η reduced PKC isoform levels by about 80% but failed to alter PHE-induced PLD activation, indicating that these PKC isoforms are not involved in PLD activation elicited by α1A-adrenergic receptor stimulation. Ectopic expression of a kinase-deficient mutant of the PKC-related kinase PKN significantly attenuated PHE-induced PLD activation. On the other hand, BIM I and Ro 31-8220 blocked PHE-mediated increase in intracellular Ca2+ but Gö 6976 and the peptide inhibitor did not. In the absence of extracellular Ca2+, PHE failed to increase PLD activity. These results indicate that α1A-adrenergic receptor-stimulated PLD activation is mediated by a mechanism independent of PKCα, δ, ε, and η, but dependent on a PKC-related kinase, PKN. Moreover, PKC inhibitors BIM I and Ro 31-8220 block PHE-induced PLD activity by inhibiting calcium signal. Caution should be used in interpreting the data obtained with PKC inhibitors in vivo.

Original languageEnglish (US)
Pages (from-to)1206-1215
Number of pages10
JournalJournal of Pharmacology and Experimental Therapeutics
Volume303
Issue number3
DOIs
StatePublished - Dec 1 2002

Fingerprint

Phospholipase D
Phenylephrine
Adrenergic Receptors
Protein Kinase C
Calcium
Protein C Inhibitor
Protein Kinase Inhibitors
Protein Isoforms
protein kinase C kinase
Peptides
Antisense Oligonucleotides
Mutant Proteins
Arachidonic Acid
Phosphotransferases

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Pharmacology

Cite this

Calcium and protein kinase C (PKC)-related kinase mediate α1A-adrenergic receptor-stimulated activation of phospholipase D in rat-1 cells, independent of PKC. / Parmentier, Jean Hugues; Ahmed, Aftab; Ruan, Ying; Gandhi, Gautam K.; Saeed, Abdelwahab E.; Malik, Kafait.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 303, No. 3, 01.12.2002, p. 1206-1215.

Research output: Contribution to journalArticle

@article{dd793b56549f4a3bbd059e33878c61f7,
title = "Calcium and protein kinase C (PKC)-related kinase mediate α1A-adrenergic receptor-stimulated activation of phospholipase D in rat-1 cells, independent of PKC",
abstract = "A previous study conducted in rat-1 cells expressing α1A-adrenergic receptors showed that phenylephrine (PHE) stimulates phospholipase D (PLD) activity. This study was conducted to determine the contribution of protein kinase C (PKC) to PHE-induced PLD activation in these cells. PKC inhibitors bisindolylmaleimide (BIM) I and Ro 31-8220, but not G{\"o} 6976 or a pseudosubstrate peptide inhibitor of PKCα, decreased PLD activity and arachidonic acid release elicited by PHE. However, antisense oligonucleotides directed against PKC α, δ, ε, and η reduced PKC isoform levels by about 80{\%} but failed to alter PHE-induced PLD activation, indicating that these PKC isoforms are not involved in PLD activation elicited by α1A-adrenergic receptor stimulation. Ectopic expression of a kinase-deficient mutant of the PKC-related kinase PKN significantly attenuated PHE-induced PLD activation. On the other hand, BIM I and Ro 31-8220 blocked PHE-mediated increase in intracellular Ca2+ but G{\"o} 6976 and the peptide inhibitor did not. In the absence of extracellular Ca2+, PHE failed to increase PLD activity. These results indicate that α1A-adrenergic receptor-stimulated PLD activation is mediated by a mechanism independent of PKCα, δ, ε, and η, but dependent on a PKC-related kinase, PKN. Moreover, PKC inhibitors BIM I and Ro 31-8220 block PHE-induced PLD activity by inhibiting calcium signal. Caution should be used in interpreting the data obtained with PKC inhibitors in vivo.",
author = "Parmentier, {Jean Hugues} and Aftab Ahmed and Ying Ruan and Gandhi, {Gautam K.} and Saeed, {Abdelwahab E.} and Kafait Malik",
year = "2002",
month = "12",
day = "1",
doi = "10.1124/jpet.102.041384",
language = "English (US)",
volume = "303",
pages = "1206--1215",
journal = "Journal of Pharmacology and Experimental Therapeutics",
issn = "0022-3565",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "3",

}

TY - JOUR

T1 - Calcium and protein kinase C (PKC)-related kinase mediate α1A-adrenergic receptor-stimulated activation of phospholipase D in rat-1 cells, independent of PKC

AU - Parmentier, Jean Hugues

AU - Ahmed, Aftab

AU - Ruan, Ying

AU - Gandhi, Gautam K.

AU - Saeed, Abdelwahab E.

AU - Malik, Kafait

PY - 2002/12/1

Y1 - 2002/12/1

N2 - A previous study conducted in rat-1 cells expressing α1A-adrenergic receptors showed that phenylephrine (PHE) stimulates phospholipase D (PLD) activity. This study was conducted to determine the contribution of protein kinase C (PKC) to PHE-induced PLD activation in these cells. PKC inhibitors bisindolylmaleimide (BIM) I and Ro 31-8220, but not Gö 6976 or a pseudosubstrate peptide inhibitor of PKCα, decreased PLD activity and arachidonic acid release elicited by PHE. However, antisense oligonucleotides directed against PKC α, δ, ε, and η reduced PKC isoform levels by about 80% but failed to alter PHE-induced PLD activation, indicating that these PKC isoforms are not involved in PLD activation elicited by α1A-adrenergic receptor stimulation. Ectopic expression of a kinase-deficient mutant of the PKC-related kinase PKN significantly attenuated PHE-induced PLD activation. On the other hand, BIM I and Ro 31-8220 blocked PHE-mediated increase in intracellular Ca2+ but Gö 6976 and the peptide inhibitor did not. In the absence of extracellular Ca2+, PHE failed to increase PLD activity. These results indicate that α1A-adrenergic receptor-stimulated PLD activation is mediated by a mechanism independent of PKCα, δ, ε, and η, but dependent on a PKC-related kinase, PKN. Moreover, PKC inhibitors BIM I and Ro 31-8220 block PHE-induced PLD activity by inhibiting calcium signal. Caution should be used in interpreting the data obtained with PKC inhibitors in vivo.

AB - A previous study conducted in rat-1 cells expressing α1A-adrenergic receptors showed that phenylephrine (PHE) stimulates phospholipase D (PLD) activity. This study was conducted to determine the contribution of protein kinase C (PKC) to PHE-induced PLD activation in these cells. PKC inhibitors bisindolylmaleimide (BIM) I and Ro 31-8220, but not Gö 6976 or a pseudosubstrate peptide inhibitor of PKCα, decreased PLD activity and arachidonic acid release elicited by PHE. However, antisense oligonucleotides directed against PKC α, δ, ε, and η reduced PKC isoform levels by about 80% but failed to alter PHE-induced PLD activation, indicating that these PKC isoforms are not involved in PLD activation elicited by α1A-adrenergic receptor stimulation. Ectopic expression of a kinase-deficient mutant of the PKC-related kinase PKN significantly attenuated PHE-induced PLD activation. On the other hand, BIM I and Ro 31-8220 blocked PHE-mediated increase in intracellular Ca2+ but Gö 6976 and the peptide inhibitor did not. In the absence of extracellular Ca2+, PHE failed to increase PLD activity. These results indicate that α1A-adrenergic receptor-stimulated PLD activation is mediated by a mechanism independent of PKCα, δ, ε, and η, but dependent on a PKC-related kinase, PKN. Moreover, PKC inhibitors BIM I and Ro 31-8220 block PHE-induced PLD activity by inhibiting calcium signal. Caution should be used in interpreting the data obtained with PKC inhibitors in vivo.

UR - http://www.scopus.com/inward/record.url?scp=0036897330&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036897330&partnerID=8YFLogxK

U2 - 10.1124/jpet.102.041384

DO - 10.1124/jpet.102.041384

M3 - Article

VL - 303

SP - 1206

EP - 1215

JO - Journal of Pharmacology and Experimental Therapeutics

JF - Journal of Pharmacology and Experimental Therapeutics

SN - 0022-3565

IS - 3

ER -