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Determination of the number of cysteine residues in high molecular weight subunits of wheat glutenin
Author(s) -
Morel MarieHélène,
Bonicel Joëlle
Publication year - 1996
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150170311
Subject(s) - glutenin , cysteine , iodoacetic acid , protein subunit , chemistry , polyacrylamide gel electrophoresis , biochemistry , molecular mass , gel electrophoresis , alkylation , chromatography , enzyme , catalysis , gene
A simple method allowing the determination of the number of cysteine residues of the high molecular weight glutenin subunits (HMW‐GS) is presented. The method was adapted from that of Creighton (T. E. Creighton, Nature 1980, 2840 , 487–489) with modified reagents for alkylation of the cysteine residues and the electrophoretic system. The acid‐urea‐polyacrylamide gel electrophoresis (PAGE) method, developed by Morel (M. H. Morel, Cereal Chem. 1994, 713 , 238–242), was adopted and mixtures of iodoacetic acid and 4‐vinylpyridine were used to alkylate the glutenin subunits. The accuracy of the method was checked with some HMW‐GS whose number of cysteine residues was already known from molecular biology approaches. In subunits 5*, 2.2, 2.2* and 4.1 (of Dx types) from cultivars Ben and Fiorello, MG 7249, MG 315, and Kador, respectively, only 4 cysteine residues were demonstrated. Subunit 20 was found to exist as a regular Bx and By subunit pair, with the Bx subunit containing only 2 cysteine residues, which is unusual.