Identification of polypeptides associated with an enriched cytoskeleton-protein body fraction from developing rice...

Recent evidence has shown that the prolamine polysomes are attached not only to the endoplasmic reticulum membranes that bound the prolamine protein bodies (PBs) but also to cytoskeleton elements associated with this subcellular fraction. To learn more about the nature of the proteins that are associated with this supra-macromolecular complex, proteins extracted from an enriched cytoskeleton-PB fraction were resolved by two-dimensional polyacrylamide gel electrophoresis under non-equilibrium conditions and analyzed for their composition by immunological and biochemical methods. Immunoblot analysis indicated the presence of the cytoskeletal proteins, actin and tubulin, and the cytoskeletal-associated protein EF1 alpha in this fraction. Microsequencing of selected polypeptides revealed a diversity of protein sequences. In addition to contaminating storage proteins which are selectively solubilized by the isolation procedure, several ribosomal proteins and histone H3 were also identified. Some of the remaining polypeptides showed partial homology to protein sequences deposited in the database, several of which are cytoskeleton-associated proteins.