Abstract
The various isoforms of the sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) are responsible for the Ca2+ uptake from the cytosol into the endoplasmic or sarcoplasmic reticulum (ER/SR). In some tissues, the activity of SERCA can be modulated by binding partners, such as phospholamban and sarcolipin. The activity of SERCA can be characterized by its apparent affinity for Ca2+ as well as maximal enzymatic velocity. Both parameters can be effectively determined by the protocol described here. Specifically, we describe the measurement of the rate of oxalate-facilitated 45Ca uptake into the SR of crude mouse ventricular homogenates. This protocol can easily be adapted for different tissues and animal models as well as cultured cells.
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Acknowledgement
This work was supported by NIH grants HL-26057 and HL-64018 to E. G. K. and AHA grant 13POST13860006 to P. A. B.
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Bidwell, P.A., Kranias, E.G. (2016). Calcium Uptake in Crude Tissue Preparation. In: Bublitz, M. (eds) P-Type ATPases. Methods in Molecular Biology, vol 1377. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3179-8_16
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DOI: https://doi.org/10.1007/978-1-4939-3179-8_16
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