Chapter twelve - soybean seed proteomics: Methods for the isolation, detection, and identification of low abundance proteins

Authors: KIM, WONSEOK - University Of Missouri; NOTT, JOEL - Iowa State University; KIM, SUNHYUNG - University Of Missouri; Krishnan, Hari

Submitted to: Methods in Enzymology
Publication Type: Book / Chapter
Publication Acceptance Date: 6/29/2022
Publication Date: 10/21/2022
Citation: Kim, W., Nott, J., Kim, S., Krishnan, H.B. 2022. Chapter twelve - soybean seed proteomics: Methods for the isolation, detection, and identification of low abundance proteins. Methods in Enzymology. 676:325-345. https://doi.org/10.1016/bs.mie.2022.07.001.
DOI: https://doi.org/10.1016/bs.mie.2022.07.001

Interpretive Summary: The two abundant seed storage proteins of soybean are the 11S and 7S globulins. In addition, soybeans also accumulate several less abundant proteins. Proteomic analysis of low abundant proteins is challenging largely due to overabundance of 7S and 11S globulins. In this study we describe simple, fast, and inexpensive methods to remove most storage proteins leading to the enrichment of nonabundant proteins within seeds. Our methodology would aid researchers in the isolation of more nonabundant seed proteins allowing for the discovery of new or novel proteins within the seed proteome, and allowing more clues about their nutritive value, yield potential, and environmental stress responses to be ascertained. A better understanding of the identity and functions of low abundant proteins will enable scientists the ability to manipulate the quality and quantity of beneficial soybean seed protein, which will benefit US soybean farmers.

Technical Abstract: The salt-soluble globulins, glycinins (11S globulin), and ß-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.