Phosphorylation of caldesmon by ERK MAP kinases in smooth muscle

Jason Hedges, Brian C. Oxhorn, Michael Carty, Leonard P. Adam, Ilia A. Yamboliev, William T. Gerthoffer

Research output: Contribution to journalArticle

85 Citations (Scopus)

Abstract

Phosphorylation of h-caldesmon has been proposed to regulate airway smooth muscle contraction. Both extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein (MAP) kinases phosphorylate h-caldesmon in vitro. To determine whether both enzymes phosphorylate caldesmon in vivo, phosphorylation-site-selective antibodies were used to assay phosphorylation of MAP kinase consensus sites. Stimulation of cultured tracheal smooth muscle cells with ACh or platelet-derived growth factor increased caldesmon phosphorylation at Ser789 by about twofold. Inhibiting ERK MAP kinase activation with 50 μM PD-98059 blocked agonist-induced caldesmon phosphorylation completely. Inhibiting p38 MAP kinases with 25 μM SB-203580 had no effect on ACh-induced caldesmon phosphorylation. Carbachol stimulation increased caldesmon phosphorylation at Ser789 in intact tracheal smooth muscle, which was blocked by the M2 antagonist AF-DX 116 (1 μM). AF-DX 116 inhibited carbachol-induced isometric contraction by 15 ± 1.4%, thus dissociating caldesmon phosphorylation from contraction. Activation of M2 receptors leads to activation of ERK MAP kinases and phosphorylation of caldesmon with little or no functional effect on isometric force. P38 MAP kinases are also activated by muscarinic agonists, but they do not phosphorylate caldesmon in vivo.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume278
Issue number4 47-4
StatePublished - 2000
Externally publishedYes

Fingerprint

Calmodulin-Binding Proteins
Phosphorylation
Extracellular Signal-Regulated MAP Kinases
Mitogen-Activated Protein Kinases
Smooth Muscle
Muscle
p38 Mitogen-Activated Protein Kinases
Chemical activation
Carbachol
Muscarinic Agonists
Isometric Contraction
Platelet-Derived Growth Factor
Muscle Contraction
Smooth Muscle Myocytes
Assays
Cells

Keywords

  • Carbachol
  • Extracellular signal-regulated kinase
  • p38 mitogen-activated protein kinase
  • PD-98059
  • SB- 203580
  • Trachea

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology
  • Physiology (medical)

Cite this

Hedges, J., Oxhorn, B. C., Carty, M., Adam, L. P., Yamboliev, I. A., & Gerthoffer, W. T. (2000). Phosphorylation of caldesmon by ERK MAP kinases in smooth muscle. American Journal of Physiology - Cell Physiology, 278(4 47-4).

Phosphorylation of caldesmon by ERK MAP kinases in smooth muscle. / Hedges, Jason; Oxhorn, Brian C.; Carty, Michael; Adam, Leonard P.; Yamboliev, Ilia A.; Gerthoffer, William T.

In: American Journal of Physiology - Cell Physiology, Vol. 278, No. 4 47-4, 2000.

Research output: Contribution to journalArticle

Hedges, J, Oxhorn, BC, Carty, M, Adam, LP, Yamboliev, IA & Gerthoffer, WT 2000, 'Phosphorylation of caldesmon by ERK MAP kinases in smooth muscle', American Journal of Physiology - Cell Physiology, vol. 278, no. 4 47-4.
Hedges J, Oxhorn BC, Carty M, Adam LP, Yamboliev IA, Gerthoffer WT. Phosphorylation of caldesmon by ERK MAP kinases in smooth muscle. American Journal of Physiology - Cell Physiology. 2000;278(4 47-4).
Hedges, Jason ; Oxhorn, Brian C. ; Carty, Michael ; Adam, Leonard P. ; Yamboliev, Ilia A. ; Gerthoffer, William T. / Phosphorylation of caldesmon by ERK MAP kinases in smooth muscle. In: American Journal of Physiology - Cell Physiology. 2000 ; Vol. 278, No. 4 47-4.
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