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Quantitative SDS–PAGE of total protein from different wheat varieties
Author(s) -
Fullington J. Garrin,
Cole Earl W.,
Kasarda Donald D.
Publication year - 1980
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.2740310108
Subject(s) - coomassie brilliant blue , chemistry , densitometry , chromatography , bradford protein assay , polyacrylamide gel electrophoresis , sodium , gel electrophoresis , electrophoresis , biochemistry , biology , staining , enzyme , genetics , physics , organic chemistry , quantum mechanics
Buffers containing sodium dodecyl sulphate (SDS) and mercaptoethanol (ME) were used to extract total protein from the flour or single seeds of five different wheat ( Triticum aestivum L.) varieties (Scout 66, Red River 68, Bankuti 1201, Atlas 66, and Omar). A dye‐binding analysis for protein was adapted to these extracts. The extracted proteins were separated according to molecular weight (mol. wt) by gel electrophoresis with SDS and the patterns were quantitated by densitometry of the gels, after proteins had been stained with Coomassie brilliant blue R250. The patterns were divided into five different areas corresponding to mol. wt ranges of 63 × 10 3 and greater (A1), 48–63 × 10 3 (A2), 40–48 × 10 3 (A3), 31–40 × 10 3 (A4), and 8–31 × 10 3 (A5). The proportions of the total area corresponding to these subareas were compared for the different varieties. Dye absorption was assumed to be directly proportional to protein concentration (in the linear range) with the same proportionality holding for the first four areas (largely storage proteins). The proteins of A5 (largely albumins) were assumed to have a three‐fold greater dye‐binding capacity than the other proteins and a correction to this area was based on this assumption. Red River 68, a variety with strong mixing characteristics, had a comparatively large proportion of its total protein in A3 and a notable peak of mol. wt 46 × 10 3 in this area. Bankuti 1201 and Omar, with weak mixing characteristics, had comparatively small proportions of their protein in A3, but large proportions in A4 (largely gliadins). Atlas 66 had a comparatively large proportion of its protein in A5 (largely albumins) and a smaller proportion in A4, but there were no strong differences in the pattern that could be related clearly to the high‐protein character of Atlas 66. All the varieties had most of their protein (65–69%) in the mol. wt ranges corresponding to A3 and A4.