Soybean seed proteomics: Methods for the isolation, detection, and identification of low abundance proteins

Abstract

The salt-soluble globulins, glycinins (11S globulin), and β-conglycinins (7S globulin), are the most abundant seed proteins of soybean seeds. Together, these two groups of proteins account for 60-70% of total soybean seed proteins. Proteomic assessment of the less abundant soybean seed proteins using general isolation protocols is challenging due to the overwhelming abundance of storage proteins. Development of a simple, fast, and inexpensive method to remove most storage proteins from a seed extract will significantly enhance the study of the nonabundant proteins within seeds. We have developed two simple methods for the depletion of abundant seed proteins resulting in the enrichment of low abundance proteins from soybean seeds. Here, we provide a detailed procedure for the isolation, separation, identification, and quantification of low abundance seed proteins of soybean.

Keywords: 2-D gel electrophoresis; Low abundant proteins; Mass spectrometry; Proteomics; Soybean; Storage proteins.