Submitted to: Journal of Basic and Applied Sciences
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: June 11, 2013
Publication Date: June 17, 2013
Citation: Natarajan, S.S., Pastor Corrales, M.A., Khan, F.H., Garrett, W.M. 2013. Proteomic analysis of common bean (Phaseolus vulgaris) by two-dimensional gel electrophoresis and mass spectrometry. Journal of Basic and Applied Sciences. 9:424-437. Interpretive Summary: Common beans are a rich and inexpensive source of proteins. They are also the most important food legume in the world especially in countries of Eastern and Southern Africa, and Latin America. Improvement of common bean seed protein is important for developing value added bean varieties. A substantial amount of information has been reported on the genetic analysis of common beans. But limited analysis of common bean protein composition is available. A better understanding of common bean protein composition is necessary to improve nutritional quality in common bean. Therefore, we have conducted studies to determine the composition of different classes of seed proteins in common beans through protein isolation and analysis. This information is very important to scientists wishing to improve the protein content in common bean.
Technical Abstract: The modern cultivated common bean (Phaseolus vulgaris) has evolved from wild common beans distributed in Central America, Mexico and the Andean region of South America. It has been reported that wild common bean accessions have higher levels of protein content than the domesticated dry bean cultivars. However, there is very little proteomic analysis of wild or domesticated common beans. Reported here is a proteomic analysis of a wild bean from Mexico conducted with the purpose of establishing a proteomic reference map for the common bean. We utilized two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) for protein separation and subsequent mass spectrometry (MS) for protein identification. Proteins were separated in the first dimension using a pH ranges from 4-7. A total of 237 protein spots were isolated, digested with trypsin, and analyzed by MALDI/TOF/TOF. We identified 141 protein spots by searching NCBI non redundant databases using the Mascot search engine and found a total of 43unique proteins. Gene Ontology (GO) analysis was employed to understand the molecular processes in which the identified common bean proteins are involved. The majority of proteins are involved in binding (41.5%) and catalytic activity (35.8%), followed by nutrient reservoir activity (7.5%), antioxidant activity (1.9%), transporter activity (3.8%), enzyme regulator activity (3.8%), structural molecule activity (1.9%), and electron carrier activity (3.8%). The results indicate that 2D-PAGE with mass spectrometry, is a sensitive and useful technique for separation and identification of common bean abundant and low abundant proteins.