The rate of activation by calmodulin of isoform 4 of the plasma membrane Ca2+ pump is slow and is changed by alternatives splicing

Ariel J. Caride, Nancy L. Elwess, Anil K. Verma, Adelaida G. Filoteo, Ágnes Enyedi, Željko Bajzer, John T. Penniston

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

A reconstitution system allowed us to measure the ATPase activity of specific isoforms of the plasma membrane Ca2+ pump continuously, and to measure the effects of adding or removing calmodulin. The rate of activation by calmodulin of isoform 4b was found to be very slow, with a half-time (at 235 nM calmodulin and 0.5 μM free Ca2+) of about 1 min. The rate of inactivation of isoform 4b when calmodulin was removed was even slower, with a half-time of about 20 min. Isoform 4a has a lower apparent affinity for calmodulin than 4b, but its activation rate was surprisingly faster (half time about 20 s). This was coupled with a much faster inactivation rate, consistent with its low affinity. A truncated mutant of isoform 4b also had a more rapid activation rate, indicating that the downstream inhibitory region of full-length 4b contributed to its slow activation. The results indicate that the slow activation is due to occlusion of the calmodulin-binding domain of 4b, caused by its strong interaction with the catalytic core. Since the activation of 4b occurs on a time scale comparable to that of many Ca2+ spikes, this phenomenon is important to the function of the pump in living cells. The slow response of 4b indicates that this isoform may be the appropriate one for cells which respond slowly to Ca2+ signals.

Original languageEnglish (US)
Pages (from-to)35227-35232
Number of pages6
JournalJournal of Biological Chemistry
Volume274
Issue number49
DOIs
StatePublished - Dec 3 1999

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Alternative Splicing
Calmodulin
Cell membranes
Protein Isoforms
Chemical activation
Cell Membrane
Pumps
Adenosine Triphosphatases
Catalytic Domain
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Caride, A. J., Elwess, N. L., Verma, A. K., Filoteo, A. G., Enyedi, Á., Bajzer, Ž., & Penniston, J. T. (1999). The rate of activation by calmodulin of isoform 4 of the plasma membrane Ca2+ pump is slow and is changed by alternatives splicing. Journal of Biological Chemistry, 274(49), 35227-35232. https://doi.org/10.1074/jbc.274.49.35227

The rate of activation by calmodulin of isoform 4 of the plasma membrane Ca2+ pump is slow and is changed by alternatives splicing. / Caride, Ariel J.; Elwess, Nancy L.; Verma, Anil K.; Filoteo, Adelaida G.; Enyedi, Ágnes; Bajzer, Željko; Penniston, John T.

In: Journal of Biological Chemistry, Vol. 274, No. 49, 03.12.1999, p. 35227-35232.

Research output: Contribution to journalArticle

Caride, AJ, Elwess, NL, Verma, AK, Filoteo, AG, Enyedi, Á, Bajzer, Ž & Penniston, JT 1999, 'The rate of activation by calmodulin of isoform 4 of the plasma membrane Ca2+ pump is slow and is changed by alternatives splicing', Journal of Biological Chemistry, vol. 274, no. 49, pp. 35227-35232. https://doi.org/10.1074/jbc.274.49.35227
Caride, Ariel J. ; Elwess, Nancy L. ; Verma, Anil K. ; Filoteo, Adelaida G. ; Enyedi, Ágnes ; Bajzer, Željko ; Penniston, John T. / The rate of activation by calmodulin of isoform 4 of the plasma membrane Ca2+ pump is slow and is changed by alternatives splicing. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 49. pp. 35227-35232.
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