The rat platelet 97-kDa Ca2+ATPase isoform is the sarcoendoplasmic reticulum Ca2+ATPase 3 protein

Régis Bobe, Raymonde Bredoux, Frank Wuytack, Rozenn Quarck, T. Kovács, Béla Papp, Elisabeth Corvazier, Clarice Magnier, Jocelyne Enouf

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Abstract

We recently showed that human and rat platelets express two types of SERCAs (Sarco Endoplasmic Reticulum Ca2+ATPases): a 100-kDa SERCA2b isoform and a 97-kDa SERCA isoform. Here, we explored the possibility that the rat 97-kDa isoform is identical to the SERCA3 protein. For this purpose, we first attempted to detect SERCA3 mRNA in rat platelet total RNA by reverse transcription-polymerase chain reaction using SERCA3-specific primers, and demonstrated the presence of this mRNA species by sequencing the amplification product. We then searched for a relationship between the expression of the SERCA3 mRNA and of the 97-kDa protein using either rat aortic smooth muscle cells, previously found not to express the 97-kDa SERCA isoform (negative model), or platelets of spontaneously hypertensive rats (SHR), which overexpress this isoform (overexpression model) but express the 100-kDa SERCA2b isoform normally. No expression of SERCA3 mRNA was detectable by analysis of smooth muscle cell RNA, but comparison by reverse transcription-polymerase chain reaction of the SERCA2b and SERCA3 mRNAs from the platelets of normotensive (Wistar-Kyoto, WKY) rats and SHR clearly demonstrated a 238 ± 43% increase in the expression of the SERCA3 mRNA in SHR platelets only. Last, by comparative Western blotting of WKY rat and SHR platelet membranes using a recently developed polyclonal anti-SERCA3 antibody, we established that the 97-kDa SERCA and the SERCA3 protein are identical, as immunostaining of the 97-kDa protein revealed a 230 ± 25% increase in the expression of this protein in SHR versus WKY rat platelets. It is concluded that the 97-kDa platelet SERCA isoform, which is up-regulated in SHR, is the SERCA3 protein. As far as we know, this constitutes the first demonstration of the actual presence of this Ca2+ATPase isoform in normal cells, in addition to the artificial transfection systems.

Original languageEnglish
Pages (from-to)1417-1424
Number of pages8
JournalJournal of Biological Chemistry
Volume269
Issue number2
Publication statusPublished - Jan 14 1994

Fingerprint

Reticulum
Calcium-Transporting ATPases
Platelets
Sarcoplasmic Reticulum Calcium-Transporting ATPases
Rats
Protein Isoforms
Inbred SHR Rats
Blood Platelets
Messenger RNA
Inbred WKY Rats
Proteins
Reverse Transcription
Smooth Muscle Myocytes
RNA
Polymerase chain reaction
Transcription
Polymerase Chain Reaction
Muscle
Cells
Transfection

ASJC Scopus subject areas

  • Biochemistry

Cite this

Bobe, R., Bredoux, R., Wuytack, F., Quarck, R., Kovács, T., Papp, B., ... Enouf, J. (1994). The rat platelet 97-kDa Ca2+ATPase isoform is the sarcoendoplasmic reticulum Ca2+ATPase 3 protein. Journal of Biological Chemistry, 269(2), 1417-1424.

The rat platelet 97-kDa Ca2+ATPase isoform is the sarcoendoplasmic reticulum Ca2+ATPase 3 protein. / Bobe, Régis; Bredoux, Raymonde; Wuytack, Frank; Quarck, Rozenn; Kovács, T.; Papp, Béla; Corvazier, Elisabeth; Magnier, Clarice; Enouf, Jocelyne.

In: Journal of Biological Chemistry, Vol. 269, No. 2, 14.01.1994, p. 1417-1424.

Research output: Contribution to journalArticle

Bobe, R, Bredoux, R, Wuytack, F, Quarck, R, Kovács, T, Papp, B, Corvazier, E, Magnier, C & Enouf, J 1994, 'The rat platelet 97-kDa Ca2+ATPase isoform is the sarcoendoplasmic reticulum Ca2+ATPase 3 protein', Journal of Biological Chemistry, vol. 269, no. 2, pp. 1417-1424.
Bobe, Régis ; Bredoux, Raymonde ; Wuytack, Frank ; Quarck, Rozenn ; Kovács, T. ; Papp, Béla ; Corvazier, Elisabeth ; Magnier, Clarice ; Enouf, Jocelyne. / The rat platelet 97-kDa Ca2+ATPase isoform is the sarcoendoplasmic reticulum Ca2+ATPase 3 protein. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 2. pp. 1417-1424.
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AU - Kovács, T.

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