|Do Carmo Silva, Anaelisabete|
Submitted to: Methods in Molecular Biology
Publication Type: Book / Chapter
Publication Acceptance Date: December 15, 2009
Publication Date: August 24, 2010
Citation: Barta, C., Do Carmo Silva, A., Salvucci, M.E., 2010. Rubisco activase activity assays. Methods in Molecular Biology. 684:375-382. Interpretive Summary: In the process of photosynthesis, plants convert sunlight into chemical energy. The energy produced by photosynthesis is then used to synthesize sugars and other foodstuffs. Previous research from this research group identified an enzyme called Rubisco activase as a major regulator of photosynthesis, particularly under heat stress. In this manuscript, we describe methods for measuring the two activities of Rubisco activase; Rubisco activation and ATP hydrolysis. By sharing our methods for assaying this enzyme with other researchers, the research in this manuscript will accelerate research on this important plant enzyme by plant biologists. Research on Rubisco activase could eventually lead to breakthroughs that improve the efficiency of photosynthesis under heat stress.
Technical Abstract: Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase functions as a mechano-chemical motor protein using the energy from ATP hydrolysis to contort the structure of its target protein, Rubisco. This action modulates the activation state of Rubisco by removing tightly-bound inhibitory sugar-phosphates from Rubisco’s catalytic sites, thereby restoring the sites to catalytic competence. This chapter reports methods developed for assaying the two activities of Rubisco activase: ATP hydrolysis and Rubisco activation.