Epidermal Growth Factor Activates m-Calpain (Calpain II), at Least in Part, by Extracellular Signal-Regulated Kinase-Mediated Phosphorylation

A. Glading, R. J. Bodnar, I. J. Reynolds, Hidenori Shiraha, L. Satish, D. A. Potter, H. C. Blair, A. Wells

Research output: Contribution to journalArticle

209 Citations (Scopus)

Abstract

How m-calpain is activated in cells has challenged investigators because in vitro activation requires near-millimolar calcium. Previously, we demonstrated that m-calpain activation by growth factors requires extracellular signal-regulated kinase (ERK); this enables tail deadhesion and allows productive motility. We now show that ERK directly phosphorylates and activates m-calpain both in vitro and in vivo. We identified serine 50 as required for epidermal growth factor (EGF)-induced calpain activation in vitro and in vivo. Replacing the serine with alanine limits activation by EGF and subsequent cell deadhesion and motility. A construct with the serine converted to glutamic acid displays constitutive activity in vivo; expression of an estrogen receptor fusion construct produces a tamoxifen-sensitive enzyme. Interestingly, EGF-induced m-calpain activation occurs in the absence of increased intracellular calcium levels; EGF triggers calpain even in the presence of intracellular calcium chelators and in calcium-free media. These data provide evidence that m-calpain can be activated through the ERK cascade via direct phosphorylation and that this activation may occur in the absence of cytosolic calcium fluxes.

Original languageEnglish
Pages (from-to)2499-2512
Number of pages14
JournalMolecular and Cellular Biology
Volume24
Issue number6
DOIs
Publication statusPublished - Mar 2004
Externally publishedYes

Fingerprint

Calpain
Extracellular Signal-Regulated MAP Kinases
Epidermal Growth Factor
Phosphorylation
Serine
Calcium
Tamoxifen
Estrogen Receptors
Alanine
Cell Movement
Glutamic Acid
Intercellular Signaling Peptides and Proteins
Research Personnel
m-calpain
Enzymes
In Vitro Techniques

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Epidermal Growth Factor Activates m-Calpain (Calpain II), at Least in Part, by Extracellular Signal-Regulated Kinase-Mediated Phosphorylation. / Glading, A.; Bodnar, R. J.; Reynolds, I. J.; Shiraha, Hidenori; Satish, L.; Potter, D. A.; Blair, H. C.; Wells, A.

In: Molecular and Cellular Biology, Vol. 24, No. 6, 03.2004, p. 2499-2512.

Research output: Contribution to journalArticle

Glading, A. ; Bodnar, R. J. ; Reynolds, I. J. ; Shiraha, Hidenori ; Satish, L. ; Potter, D. A. ; Blair, H. C. ; Wells, A. / Epidermal Growth Factor Activates m-Calpain (Calpain II), at Least in Part, by Extracellular Signal-Regulated Kinase-Mediated Phosphorylation. In: Molecular and Cellular Biology. 2004 ; Vol. 24, No. 6. pp. 2499-2512.
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