Correlated expression of the 97 kDa sarcoendoplasmic reticulum Ca2+-ATPase and Rap1B in platelets and various cell lines

C. Magnier, R. Bredoux, T. Kovács, R. Quarck, B. Papp, E. Corvazier, J. De Gunzburg, J. Enouf

Research output: Contribution to journalArticle

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Abstract

Evidence has accumulated that cyclic AMP (cAMP)-induced phosphorylation of a Ras-related protein (Rap1) regulates platelet Ga2+ transport. As this transport was recently found to be controlled by two isoforms of sarcoendoplasmic reticulum Ca2+-ATPase (SERCA), the 100 kDa SERCA2b and the newly identified 97 kDa SERCA, we attempted to establish which isoform is involved in this regulation. For this purpose, we studied the expression and regulation of both the SERCA and Rap1 isoforms in platelets, haemopoietic cells and various cancer cell lines. SERCA2b was shown to be equally expressed in all the cell lines tested, as determined by detection of its phosphoenzyme formation and by Western blotting using an isoform-specific antibody. In contrast, the expression of the 97 kDa SERCA, studied by the same methods, varied from total absence in the cancer cells to high levels in the megakaryocytic cell lines. With regard to the potential regulatory Rap1 proteins, Western blotting showed different expression of total Rap1 isoforms among the cell lineages, thus ruling out any possible relationship between Rap1 and SERCA2b. However, the expression of Rap1 proteins correlated with that of the 97 kDa SERCA isoform. More refined analysis of the rap1A and rap1B isoforms by reverse transcription PCR and by determining cAMP-induced phosphorylation of Rap1B, i.e. its functional mechanism, confirmed the correlation between Rap1B and the 97 kDa SERCA expression. This relationship was also established by the concerted up-regulation of these two proteins demonstrated in the pathological model of platelets from hypertensive rats. It is concluded that the expressions of 97 kDa SERCA and Rap1B are related, suggesting that regulation of the platelet Ca2+-ATPase system by cAMP-induced phosphorylation of Rap1B specifically involves the 97 kDa SERCA.

Original languageEnglish
Pages (from-to)343-350
Number of pages8
JournalBiochemical Journal
Volume297
Issue number2
Publication statusPublished - 1994

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Reticulum
Calcium-Transporting ATPases
Platelets
Blood Platelets
Cells
Cell Line
Protein Isoforms
Phosphorylation
Cyclic AMP
Proteins
Western Blotting
ras Proteins
Cell Lineage
Transcription
Reverse Transcription
Rats
Neoplasms
Up-Regulation
Polymerase Chain Reaction
Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

Magnier, C., Bredoux, R., Kovács, T., Quarck, R., Papp, B., Corvazier, E., ... Enouf, J. (1994). Correlated expression of the 97 kDa sarcoendoplasmic reticulum Ca2+-ATPase and Rap1B in platelets and various cell lines. Biochemical Journal, 297(2), 343-350.

Correlated expression of the 97 kDa sarcoendoplasmic reticulum Ca2+-ATPase and Rap1B in platelets and various cell lines. / Magnier, C.; Bredoux, R.; Kovács, T.; Quarck, R.; Papp, B.; Corvazier, E.; De Gunzburg, J.; Enouf, J.

In: Biochemical Journal, Vol. 297, No. 2, 1994, p. 343-350.

Research output: Contribution to journalArticle

Magnier, C, Bredoux, R, Kovács, T, Quarck, R, Papp, B, Corvazier, E, De Gunzburg, J & Enouf, J 1994, 'Correlated expression of the 97 kDa sarcoendoplasmic reticulum Ca2+-ATPase and Rap1B in platelets and various cell lines', Biochemical Journal, vol. 297, no. 2, pp. 343-350.
Magnier, C. ; Bredoux, R. ; Kovács, T. ; Quarck, R. ; Papp, B. ; Corvazier, E. ; De Gunzburg, J. ; Enouf, J. / Correlated expression of the 97 kDa sarcoendoplasmic reticulum Ca2+-ATPase and Rap1B in platelets and various cell lines. In: Biochemical Journal. 1994 ; Vol. 297, No. 2. pp. 343-350.
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