Polypeptide compositions and antigenic homologies among prolamins from Italian, common and Japanese millet cultivars

Prolamins from Italian, common and Japanese millet cultivars were separately extracted with three kinds of alcohol (ethanol, propanol and isopropanol) and their component polypeptides and immunochemical relationships were examined by SDS-PAGE, two-dimensional electrophoresis and immunoblot analysis using the antiserum raised against Italian millet prolamin. Polypeptides of the individual millet prolamins appeared to be primarily composed of two major common subunit complexes with respective molecular masses ranging from about 19 to 23 kDa and from 13 to 14 kDa (groups A and B, respectively), although a few minor variations due to varietal differences were seen. Group A clearly contained one neutral subunit (21 kDa) and one basic one (22 kDa), while group B had one basic 14 kDa subunit. The Italian millet prolamin antiserum strongly bound with all three prolamin polypeptides but neither with the homologous non-prolamin fractions nor with the heterogeneous prolamins of the Triticeae (wheat, barley and rye). Amino acid compositions were found to be almost identical to one another.