|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. Purification of rubisco activase from leaves or after expression in escherichia coli. Methods in Molecular Biology. 684:363-374. 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 the purification of Rubisco activase from leaves. In addition, we show how to purify the Rubisco activase enzyme from the bacterium, E. coli, after engineering the plant gene. By sharing our methods with other researchers, the methods described in this manuscript will accelerate research on this important plant enzyme in plant biology. Research on Rubisco activase could eventually lead to breakthroughs that improve the efficiency of photosynthesis under heat stress.
Technical Abstract: Rubisco activase is a molecular chaperone that modulates the activation state of Rubisco by catalyzing the ATP-dependent removal of tightly-bound inhibitory sugar-phosphates from Rubisco’s catalytic sites. This chapter reports methods developed for the purification of native and recombinant Rubisco activase from leaves and bacterial cells, respectively.