Quantification of Rubisco activase content in leaf extracts

Rubisco activase functions to promote and maintain the catalytic activity of Rubisco. Studies with the activase-lacking Arabidopsis rca mutant (Salvucci et al. Photosynth Res 7:193-201, 1985; Salvucci et al. Plant Physiol 80:655-659, 1986), antisense activase tobacco, Arabidopsis and Flaveria bidentis plants (Mate et al. Plant Physiol 102:1119-1128, 1993; Eckardt et al. Plant Physiol 113:575-586, 1997; von Caemmerer et al. Plant Physiol 137:747-755, 2005) have shown that photosynthesis at atmospheric levels of CO2 is severely impaired when plants lack activase because Rubisco becomes sequestered in an inactive form. Activase protein has been detected in all plant species, including C3 and C4 plants and green algae (Salvucci et al. Plant Physiol 84:930-936, 1987). Rubisco activase is essential in all these photosynthetic organisms for photosynthesis and plant growth. The physiological importance of Rubisco activase is reinforced by recent studies indicating that it plays a role in the response of photosynthesis to temperature. In this chapter, we describe how to extract and quantify Rubisco activase content in leaf