Assessment of the extracellular and intracellular actions of sphingosine 1-phosphate by using the p42/p44 mitogen-activated protein kinase cascade as a model

D Tolan, A M Conway, S Rakhit, N Pyne, S Pyne

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

We have investigated the extracellular and intracellular actions of sphingosine 1-phosphate (S1P) by using cultured airway smooth muscle cells. We have demonstrated that exogenous S1P elicited an activation of mitogen activated protein kinase (p42/p44 MAPK) that was abolished by pertussis toxin (0.1 mu g/mL, 24 h), which was used to inactivate G(i). The effect of exogenous S1P might therefore be attributed to an action at a putative Gi-coupled receptor. The regulation of the p42/p44 MAPK cascade by S1P was also shown to include a protein kinase C (PKC)-dependent intermediate step. Platelet-derived growth factor (PDGF) stimulates intracellular S1P formation and was therefore used to evaluate the intracellular action of S1P. This has previously been investigated by others using the sphingosine kinase inhibitors D,L-threo-dihydrosphingosine and N,N-dimethylsphingosine. We have demonstrated here that both inhibitors block the PDGF-dependent activation of p42/p44 MAPK. However, both are also PKC inhibitors, which might account for their effect. because PDGF utilises PKC as an intermediate in the regulation of the p42/p44 MAPK cascade. Significantly, sphingosine, which is the substrate of sphingosine kinase and a PKC inhibitor, blocked the activation of p42/p44 MAPK by PDGF with an almost identical concentration dependence compared with D,L-threo-dihydrosphingosine and N,N-dimethylsphingosine. Therefore, the use of so-called sphingosine kinase inhibitors might lead to misleading interpretations because of their additional effect on PKC. Other approaches, such as oligodeoxynucleotide anti-sense against sphingosine kinase, are required to address the intracellular role of S1P. CELL SIGNAL 11;5:349-354, 1999.

LanguageEnglish
Pages349-354
Number of pages6
JournalCellular Signalling
Volume11
Issue number5
DOIs
Publication statusPublished - May 1999

Fingerprint

Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3
Protein Kinase C
Platelet-Derived Growth Factor
Protein C Inhibitor
Protein Kinase Inhibitors
Sphingosine
Oligodeoxyribonucleotides
Pertussis Toxin
sphingosine 1-phosphate
Mitogen-Activated Protein Kinases
Smooth Muscle Myocytes
sphingosine kinase

Keywords

  • mitogen-activated protein kinase
  • sphingosine 1-phosphate
  • PDGF
  • smooth muscle
  • D,L-threo-dihydrosphingosine
  • signal transduction pathways
  • sphingomyelin-derived lipids
  • airway smooth muscle
  • phospholipase-D activation
  • regulated kinase-2
  • membrane receptor
  • high affinity
  • sphingosine-1-phosphate
  • sphingolipids

Cite this

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title = "Assessment of the extracellular and intracellular actions of sphingosine 1-phosphate by using the p42/p44 mitogen-activated protein kinase cascade as a model",
abstract = "We have investigated the extracellular and intracellular actions of sphingosine 1-phosphate (S1P) by using cultured airway smooth muscle cells. We have demonstrated that exogenous S1P elicited an activation of mitogen activated protein kinase (p42/p44 MAPK) that was abolished by pertussis toxin (0.1 mu g/mL, 24 h), which was used to inactivate G(i). The effect of exogenous S1P might therefore be attributed to an action at a putative Gi-coupled receptor. The regulation of the p42/p44 MAPK cascade by S1P was also shown to include a protein kinase C (PKC)-dependent intermediate step. Platelet-derived growth factor (PDGF) stimulates intracellular S1P formation and was therefore used to evaluate the intracellular action of S1P. This has previously been investigated by others using the sphingosine kinase inhibitors D,L-threo-dihydrosphingosine and N,N-dimethylsphingosine. We have demonstrated here that both inhibitors block the PDGF-dependent activation of p42/p44 MAPK. However, both are also PKC inhibitors, which might account for their effect. because PDGF utilises PKC as an intermediate in the regulation of the p42/p44 MAPK cascade. Significantly, sphingosine, which is the substrate of sphingosine kinase and a PKC inhibitor, blocked the activation of p42/p44 MAPK by PDGF with an almost identical concentration dependence compared with D,L-threo-dihydrosphingosine and N,N-dimethylsphingosine. Therefore, the use of so-called sphingosine kinase inhibitors might lead to misleading interpretations because of their additional effect on PKC. Other approaches, such as oligodeoxynucleotide anti-sense against sphingosine kinase, are required to address the intracellular role of S1P. CELL SIGNAL 11;5:349-354, 1999.",
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Assessment of the extracellular and intracellular actions of sphingosine 1-phosphate by using the p42/p44 mitogen-activated protein kinase cascade as a model. / Tolan, D ; Conway, A M ; Rakhit, S ; Pyne, N ; Pyne, S .

In: Cellular Signalling, Vol. 11, No. 5, 05.1999, p. 349-354.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Assessment of the extracellular and intracellular actions of sphingosine 1-phosphate by using the p42/p44 mitogen-activated protein kinase cascade as a model

AU - Tolan, D

AU - Conway, A M

AU - Rakhit, S

AU - Pyne, N

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N2 - We have investigated the extracellular and intracellular actions of sphingosine 1-phosphate (S1P) by using cultured airway smooth muscle cells. We have demonstrated that exogenous S1P elicited an activation of mitogen activated protein kinase (p42/p44 MAPK) that was abolished by pertussis toxin (0.1 mu g/mL, 24 h), which was used to inactivate G(i). The effect of exogenous S1P might therefore be attributed to an action at a putative Gi-coupled receptor. The regulation of the p42/p44 MAPK cascade by S1P was also shown to include a protein kinase C (PKC)-dependent intermediate step. Platelet-derived growth factor (PDGF) stimulates intracellular S1P formation and was therefore used to evaluate the intracellular action of S1P. This has previously been investigated by others using the sphingosine kinase inhibitors D,L-threo-dihydrosphingosine and N,N-dimethylsphingosine. We have demonstrated here that both inhibitors block the PDGF-dependent activation of p42/p44 MAPK. However, both are also PKC inhibitors, which might account for their effect. because PDGF utilises PKC as an intermediate in the regulation of the p42/p44 MAPK cascade. Significantly, sphingosine, which is the substrate of sphingosine kinase and a PKC inhibitor, blocked the activation of p42/p44 MAPK by PDGF with an almost identical concentration dependence compared with D,L-threo-dihydrosphingosine and N,N-dimethylsphingosine. Therefore, the use of so-called sphingosine kinase inhibitors might lead to misleading interpretations because of their additional effect on PKC. Other approaches, such as oligodeoxynucleotide anti-sense against sphingosine kinase, are required to address the intracellular role of S1P. CELL SIGNAL 11;5:349-354, 1999.

AB - We have investigated the extracellular and intracellular actions of sphingosine 1-phosphate (S1P) by using cultured airway smooth muscle cells. We have demonstrated that exogenous S1P elicited an activation of mitogen activated protein kinase (p42/p44 MAPK) that was abolished by pertussis toxin (0.1 mu g/mL, 24 h), which was used to inactivate G(i). The effect of exogenous S1P might therefore be attributed to an action at a putative Gi-coupled receptor. The regulation of the p42/p44 MAPK cascade by S1P was also shown to include a protein kinase C (PKC)-dependent intermediate step. Platelet-derived growth factor (PDGF) stimulates intracellular S1P formation and was therefore used to evaluate the intracellular action of S1P. This has previously been investigated by others using the sphingosine kinase inhibitors D,L-threo-dihydrosphingosine and N,N-dimethylsphingosine. We have demonstrated here that both inhibitors block the PDGF-dependent activation of p42/p44 MAPK. However, both are also PKC inhibitors, which might account for their effect. because PDGF utilises PKC as an intermediate in the regulation of the p42/p44 MAPK cascade. Significantly, sphingosine, which is the substrate of sphingosine kinase and a PKC inhibitor, blocked the activation of p42/p44 MAPK by PDGF with an almost identical concentration dependence compared with D,L-threo-dihydrosphingosine and N,N-dimethylsphingosine. Therefore, the use of so-called sphingosine kinase inhibitors might lead to misleading interpretations because of their additional effect on PKC. Other approaches, such as oligodeoxynucleotide anti-sense against sphingosine kinase, are required to address the intracellular role of S1P. CELL SIGNAL 11;5:349-354, 1999.

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KW - smooth muscle

KW - D,L-threo-dihydrosphingosine

KW - signal transduction pathways

KW - sphingomyelin-derived lipids

KW - airway smooth muscle

KW - phospholipase-D activation

KW - regulated kinase-2

KW - membrane receptor

KW - high affinity

KW - sphingosine-1-phosphate

KW - sphingolipids

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