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| Content Provider | Journal of Biological Chemistry (JBC) |
|---|---|
| Author | Robert, Valerie Giorgi, Francesca De Massimino, Maria Lina Cantini, Marcello Pozzan, Tullio |
| Abstract | Direct monitoring of the free Ca2+ concentration in the sarcoplasmic reticulum (SR) was carried out in rat skeletal myotubes transfected with a specifically targeted aequorin chimera (srAEQ). Myotubes were also transfected with a chimeric aequorin (erAEQ) that we have demonstrated previously is retained in the endoplasmic reticulum (ER). Immunolocalization analysis showed that although both recombinant proteins are distributed in an endomembrane network identifiable with immature SR, the erAEQ protein was retained also in the perinuclear membrane. The difficulty of measuring [Ca2+] in 100–1000 μm range was overcome with the use of the synthetic coelenterazine analogue, coelenterazine n. We demonstrate that the steady state levels of [Ca2+] measured with srAEQ is around 300 μm, whereas that measured with erAEQ is significantly lower, i.e. around 200 μm. The effects of caffeine, high KCl, and nicotinic receptor stimulation, in the presence or absence of external calcium or after blockade of the Ca-ATPase, were investigated with both chimeras. The kinetics of [Ca2+] changes revealed by the erAEQ were similar, but not identical, neither quantitatively nor qualitatively, to those monitored with the srAEQ, indicating that at this stage of muscle development, differences exist between SR and ER in their mechanisms of Ca2+ handling. The functional implications of these findings are discussed. |
| Related Links | http://www.jbc.org/content/273/46/30372.abstract |
| Ending Page | 30378 |
| Starting Page | 30372 |
| Page Count | 7 |
| File Format | HTM / HTML PDF |
| ISSN | 00219258 |
| Journal | Journal of Biological Chemistry (JBC) |
| Issue Number | 46 |
| Volume Number | 273 |
| DOI | 10.1074/jbc.273.46.30372 |
| e-ISSN | 1083351X |
| Language | English |
| Publisher | American Society for Biochemistry and Molecular Biology |
| Publisher Date | 1998-11-13 |
| Access Restriction | Open |
| Subject Keyword | Sarcoplasmic reticulum (SR) Endoplasmic reticulum (ER) Tert-butylhydroquinone (tBuBHQ) Dihydropyridine receptor (DHPR) Ryanodine receptor (RyR) Sarco-endoplasmic reticulum Ca2+ATPase. (SERCA) CELL BIOLOGY AND METABOLISM |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology |
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