Poster: Protein turnover

Abs # 813: Protein dynamics during soybean seed development

Presenter:	Miernyk, Jan A, miernykj@missouri.edu
Authors	Miernyk, Jan A (A) (B)  Herman, Eliot M (A) (D)  Mooney, Brian P (B) (C)
Affiliations:	(A): USDA, ARS, Plant Genetics Research Unit (B): Department of Biochemistry, University of Missouri, Columbia, MO (C): Proteomics Center, University of Missouri, Columbia, MO (D): The Donald Danforth Plant Science Center, St. Louis, MO

Embryos were harvested at specific stages of soybean (Glycine max L. cv. Jack) seed development. Total protein fractions were isolated from seeds at each stage, and the components separated by 2-dimensional electrophoresis. Sequential sets of proteins were stained with CyeDye3 and CyeDye5 for difference gel electrophoresis (DIGE) analysis. When these gels were subject to image analysis, proteins present at a constant level appeared as yellow spots, proteins that increased in abundance appeared green, and proteins that decreased in abundance appeared red. Selected proteins were subjected to in gel tryptic digestion, and peptide mass fingerprint analysis by MALDI-TOF mass spectrometry. At the later stages of seed development, multiplex analysis was conducted where gels were stained for glycoproteins with Pro-Q Emerald, or phosphoproteins with Pro-Q Diamond, and then for total proteins with Coomassie Blue. Combining these methods, we hope to prepare a catalog of protein changes during oilseed development, and to use this as the basis for analysis of the mechanisms that regulate protein re-modeling in plant cells.