Disulphide bonds in wheat gluten: further cystine peptides from high molecular weight (HMW) and low molecular weight (LMW) subunits of glutenin and from gamma-gliadins.
Glutenin was prepared from gluten of the wheat variety Rektor by extraction of gliadin with aqueous ethanol. It was cleaved successively into soluble peptides by the enzymes trypsin and thermolysin. Separation of the peptide mixtures was performed by gel permeation chromatography (GPC) on Sephadex G25 and reversed phase high performance liquid chromatography (RP-HPLC) on ODS-Hypersil. Cystine peptides were detected by differential chromatography of the samples prior to and after reduction. After isolation by multi-step RP-HPLC, the cystine peptides were reduced. The resulting cysteine peptides were alkylated with 4-vinylpyridine, separated by RP-HPLC and sequenced by means of the Edman degradation. The isolated cystine peptides represented a considerable portion of the total cysteine in glutenin: four out of seven cysteine residues of HMW subunits, and eight out of nine cysteine residues of LMW subunits are documented by at least one cystine peptide. Most of the peptides corresponded to known sequences of gluten protein components. From the structures of some tryptic peptides, inter- and intramolecular disulphide bonds for HMW subunits of glutenin have been proven. Cystine peptides from the thermolytic digest have been assigned to LMW subunits of glutenin and to gamma-gliadins. Other peptides have been closely related to partial sequences of these protein components. The results have allowed several conclusions about the arrangement of intra- and intermolecular disulphide bridges in gluten proteins.
Köhler
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P
Deutsche Forschungsanstalt für Lebensmittelchemie, München, Germany.
Belitz
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HD
Wieser
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H
D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003553:Cystine / Q000032:analysis*; D004220:Disulfides; D005903:Gliadin / Q000737:chemistry*; D005983:Glutens / Q000031:analogs & derivatives* / Q000737:chemistry*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D008970:Molecular Weight; D013820:Thermolysin; D014908:Triticum; D014357:Trypsin
D004220:Disulfides; D003553:Cystine; D005983:Glutens; D005903:Gliadin; D014357:Trypsin; D013820:Thermolysin; C037302:glutenin