Abstract
It has recently been reported that tomato fruit chromoplasts can synthesize ATP de novo using an ATP synthase complex harboring an atypical γ-subunit which is also present in a variety of plant species. However many aspects related with the biochemical processes underlying this process remain largely unknown. Here we describe detailed protocols for the isolation of tomato fruit chromoplasts and the determination of ATP levels (end-point measurements) and ATP synthesis rates (kinetic measurements) in these organelles using bioluminescent luciferin/luciferase based assays.
Keywords: Chromoplast, Tomato, Fruit, ATP levels
Materials and Reagents
Equipment
Software
Procedure
Recipes
Acknowledgments
This protocol has been adapted from methods previously described in Angaman et al. (2012) and Pateraki et al. (2013). This work has been supported by grants from the Spanish Ministerio de Ciencia e Innovación (BIO2009-09523) and Ministerio de Economía y Competitividad (AGL2013-43522-R), both including FEDER Funds, the Spanish Consolider-Ingenio 2010 Program (CSD2007-00036 Centre for Research in Agrigenomics) and the Generalitat de Catalunya (2009SGR0026). Marta Renato is a recipient of a predoctoral fellowship from the Spanish Ministerio de Educación, Cultura y Deporte.
References
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