Abstract
The plasma membrane calcium–calmodulin-dependent ATPase (PMCA) is a calcium-extruding enzymatic pump that ejects calcium from the cytoplasm to the extracellular compartment. Although in excitable cells such as skeletal and cardiac muscle cells PMCA has been shown to play only a minor role in regulating global intracellular calcium concentration, increasing evidence points to an important role for PMCA in signal transduction, in particular in the nitric oxide signaling pathway. Moreover, recent evidence has shown the functional importance of PMCA in mediating cardiac contractility and vascular tone. Here we describe a method in determining PMCA activity in the microsomal membrane preparation from cultured cells that overexpress specific isoform of PMCA by using modified coupled enzyme assay.
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Acknowledgments
This work was supported by Medical Research Council (MRC) Programme Grant (G0500025) to L.N. and British Heart Foundation (BHF) Project Grant (PG/05/082) to D.O and E.C. The laboratory is also supported by the NIHR Biomedical Research Funding Scheme. TM is a lecturer in Biochemistry Dept., Zagazig University, Egypt. Address for correspondence to: Dr Delvac Oceandy (http://delvac.oceandy@manchester.ac.uk).
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Mohamed, T.M.A., Baudoin-Stanley, F.M., Abou-Leisa, R., Cartwright, E., Neyses, L., Oceandy, D. (2010). Measurement of Plasma Membrane Calcium–Calmodulin-Dependent ATPase (PMCA) Activity. In: Yan, Q. (eds) Membrane Transporters in Drug Discovery and Development. Methods in Molecular Biology, vol 637. Humana Press. https://doi.org/10.1007/978-1-60761-700-6_18
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DOI: https://doi.org/10.1007/978-1-60761-700-6_18
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