|Hurkman Ii, William|
Submitted to: International Gluten Workshop
Publication Type: Proceedings
Publication Acceptance Date: September 7, 2009
Publication Date: September 1, 2010
Citation: Dupont, F.M., Altenbach, S.B., Vensel, W.H., Tanaka, C.K., Hurkman II, W.J. 2010. Differential effects of mineral nutrition on wheat flour proteins determined by quantitative 2-DE and MS/MS. International Gluten Workshop. p.64-67. Technical Abstract: Flour protein composition changed significantly when plants of the hard red spring wheat Triticum aestivum Butte 86 were grown with or without post-anthesis fertilization with NPK. To measure changes in individual proteins, 369 protein spots were resolved and quantified by 2-DE of total flour protein extracts. Of these, 188 spots were identified by MS/MS, accounting for 88 to 91% of total protein. To identify and distinguish among the gliadins and low molecular weight glutenin subunits (LMW-GS), it was necessary to use 3 proteases and an improved protein sequence database that included gliadin and LMW-GS sequences translated from DNA contigs assembled from Butte 86 ESTs. As previously reported, percentages of S-rich protein types decreased and S-poor protein types increased when high protein flour was obtained by use of post-anthesis NPK. Components of the glutenin polymer changed greatly. High molecular weight-glutenin subunits (HMW-GS) increased and LMW-GS decreased as a percent of total flour protein, such that the ratio of LMW-GS to HMW-GS decreased from approximately 1.4 to 0.8. Alpha-gliadins increased from 14 to 17% and gamma-gliadins decreased from 9 to 8%. Putative omega-gliadin spots increased from 4 to 9%. Additional spots with more than one gliadin type comprised around 10% of total protein. Alpha-amylase and protease inhibitors decreased from 9 to 4%, beta-amylases increased from 1.1 to 1.6%, and other enzymes decreased from 1.8 to 1.3%. Serpins increased from 1.1 to 1.5%, and triticins remained at 1.6 to 1.7%. Tritin was only 0.1% of total.