Modulation of erythrocyte Ca2+-ATPase by selective calpain cleavage of the calmodulin-binding domain

P. James, T. Vorherr, J. Krebs, A. Morelli, G. Castello, Daniel J Mc Cormick, J. T. Penniston, A. De Flora, E. Carafoli

Research output: Contribution to journalArticle

111 Citations (Scopus)

Abstract

The activity of the membrane-bound and the purified erythrocyte Ca2+-ATPase in the absence of calmodulin was stimulated by calpain digestion but could be further increased to maximal levels by calmodulin (CaM). Thus, CaM sensitivity was retained by the digested ATPase, at least at short times of incubation. In membranes digested at higher temperatures and in the purified ATPase digested at higher calpain/ATPase ratios, the ATPase became fully activated. The membrane-bound and the purified 138-kDa ATPase were converted by calpain to a fragment of approximately 124 kDa which still bound CaM and could be isolated on CaM columns when proteolysis occurred slowly by not when it occurred rapidly. Carboxypeptidase digestion of the purified enzyme and of its fragment of about 124 kDa has shown that calpain attacked the CaM-binding domain near the C terminus of the ATPase. This has also been supported by digestion of the purified enzyme and of its fragment of about 124 kDa. A first cut occurred in the middle of the domain producing a fragment of about 14 kDa and a (CaM-binding) fragment of about 124 kDa. A second cut closer to the N terminus of the domain also prodcued a fragment of about 124 kDa and accounted for the loss of CaM binding at prolonged times of incubation of the ATPase with calpain.

Original languageEnglish (US)
Pages (from-to)8289-8296
Number of pages8
JournalJournal of Biological Chemistry
Volume264
Issue number14
StatePublished - 1989
Externally publishedYes

Fingerprint

Calpain
Calcium-Transporting ATPases
Calmodulin
Adenosine Triphosphatases
Erythrocytes
Modulation
Digestion
Membranes
Proteolysis
Carboxypeptidases
Enzymes
Temperature

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

James, P., Vorherr, T., Krebs, J., Morelli, A., Castello, G., Mc Cormick, D. J., ... Carafoli, E. (1989). Modulation of erythrocyte Ca2+-ATPase by selective calpain cleavage of the calmodulin-binding domain. Journal of Biological Chemistry, 264(14), 8289-8296.

Modulation of erythrocyte Ca2+-ATPase by selective calpain cleavage of the calmodulin-binding domain. / James, P.; Vorherr, T.; Krebs, J.; Morelli, A.; Castello, G.; Mc Cormick, Daniel J; Penniston, J. T.; De Flora, A.; Carafoli, E.

In: Journal of Biological Chemistry, Vol. 264, No. 14, 1989, p. 8289-8296.

Research output: Contribution to journalArticle

James, P, Vorherr, T, Krebs, J, Morelli, A, Castello, G, Mc Cormick, DJ, Penniston, JT, De Flora, A & Carafoli, E 1989, 'Modulation of erythrocyte Ca2+-ATPase by selective calpain cleavage of the calmodulin-binding domain', Journal of Biological Chemistry, vol. 264, no. 14, pp. 8289-8296.
James, P. ; Vorherr, T. ; Krebs, J. ; Morelli, A. ; Castello, G. ; Mc Cormick, Daniel J ; Penniston, J. T. ; De Flora, A. ; Carafoli, E. / Modulation of erythrocyte Ca2+-ATPase by selective calpain cleavage of the calmodulin-binding domain. In: Journal of Biological Chemistry. 1989 ; Vol. 264, No. 14. pp. 8289-8296.
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