New methods for extraction and quantitation of zeins reveal a high content of gamma-zein in modified opaque-2 maize

We have developed methods for quantitative extraction and analysis of zeins from maize (Zea mays L.) flour. Extraction involved solubilization of total endosperm proteins in an alkaline buffer containing SDS and 2-mercaptoethanol with subsequent precipitation of nonzein proteins by the addition of e...

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Bibliographic Details
Published in:Plant physiology (Bethesda) 1990-01, Vol.92 (1), p.191-196
Main Authors: Wallace, J.C. (Bucknell University, Lewisburg, PA), Lopes, M.A, Paiva, E, Larkins, B.A
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Antibodies
Antigens
Biological and medical sciences
COMPOSICION
COMPOSITION
CONTENIDO PROTEICO
Corn
Endosperm
Enzyme linked immunosorbent assay
Ethanol
EXTRACCION
EXTRACTION
Flint
Fundamental and applied biological sciences. Psychology
Genotypes
GRAINE
INDIANA
Miscellaneous
Phenotypes
PROLAMINAS
PROLAMINE
PROTEINAS
PROTEINE
Proteins
SEMILLA
Solubilization
TENEUR EN PROTEINES
ZEA MAYS
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Summary:We have developed methods for quantitative extraction and analysis of zeins from maize (Zea mays L.) flour. Extraction involved solubilization of total endosperm proteins in an alkaline buffer containing SDS and 2-mercaptoethanol with subsequent precipitation of nonzein proteins by the addition of ethanol to 70%. Analysis of these proteins by SDS-PAGE with Coomassie blue staining and by Western blotting and ELISA assay with zein antibodies revealed that this extraction method is more quantitative than the traditional Landry-Moureaux procedure, especially for the beta- and gamma-zeins. This method was used to extract and analyze the zein content of several 'Quality Protein Maize' (QPM) varieties developed by the International Maize and Wheat Improvement Center. QPM varieties contain 'modifier genes' that confer a vitreous phenotype on opaque-2 genotypes, while maintaining the elevated levels of lysine and tryptophan characteristic of this mutant. This analysis revealed that the QPM types contain 2 to 4 times the amount of the gamma-zein than unmodified opaque-2 or normal maize varieties. Possible relationships between the high expression of the gamma-zein and the modified opaque phenotype are discussed
ISSN:0032-0889
1532-2548
DOI:10.1104/pp.92.1.191