Enrichment of the Plant Cytosolic Fraction
The cytosol is at the core of cellular metabolism and contains many important metabolic pathways, including glycolysis, gluconeogenesis, and the pentose phosphate pathway. Despite the importance of this matrix, few attempts have sought to specifically enrich this compartment from plants. Although a variety of biochemical pathways and signaling cascades pass through the cytosol, much of the focus has usually been targeted at the reactions that occur within membrane-bound organelles of the plant cell. In this chapter, we outline a method for the enrichment of the cytosol from rice suspension cell cultures which includes sample preparation and enrichment as well as validation using immunoblotting and fluorescence-tagged proteins.
Key wordsCytosol Cytoplasm Rice Cell culture
This work was supported by the U. S. Department of Energy, Office of Science, Office of Biological and Environmental Research, through contract [DE-AC02-05CH11231] between Lawrence Berkeley National Laboratory and the U. S. Department of Energy. JLH is supported by an Australian Research Council (ARC) Future Fellowship [FT130101165].