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Aequorin Luminescence-Based Functional Calcium Assay for Heterotrimeric G-Proteins in Arabidopsis

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Heterotrimeric GTP-binding proteins (G-proteins) and G-protein-coupled receptors are important signaling components in eukaryotes. In plants, the G-proteins are involved in diverse physiological processes, some of which are exerted via changes in the level of cytosolic free calcium concentration ([Ca2+ ]cyt ). Various techniques have been developed to measure the change of [Ca2+ ]cyt , e.g., calcium-sensitive microelectrodes, chemical fluorescent dyes, and biosensors based on luminescent or fluorescent indicators. In this chapter, we describe a protocol for in vivo [Ca2+ ]cyt measurement in G-protein mutants expressing aequorin, a luminescent-based calcium biosensor, to extend our knowledge about G-protein mediated Ca2+ signaling. This method is also applicable to other early signaling events that are mediated by changes in [Ca2+ ]cyt levels.
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