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Investigation and correction of the gene‐derived sequence of glutenin subunit 1Dx2 by matrix‐assisted laser desorption/ionisation mass spectrometry
Author(s) -
Cunsolo Vincenzo,
Foti Salvatore,
Saletti Rosaria,
Gilbert Simon,
Tatham Arthur S.,
Shewry Peter R.
Publication year - 2002
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.807
Subject(s) - chemistry , glutenin , mass spectrometry , chromatography , protein mass spectrometry , protein subunit , matrix assisted laser desorption/ionization , peptide sequence , molecular mass , matrix (chemical analysis) , analytical chemistry (journal) , desorption , gene , biochemistry , electrospray ionization , organic chemistry , adsorption , enzyme
Direct matrix‐assisted laser desorption/ionisation time‐of‐flight mass spectrometry (MALDI‐TOFMS) analysis of a mixture of tryptic peptides was used to verify the gene‐derived amino acid sequence of the high molecular weight (HMW) subunit 1Dx2 of bread wheat. Analysis of the digest was performed by recording several MALDI mass spectra of the mixture at low, medium and high mass ranges, and optimising the matrix and the acquisition parameters for each mass range. This resulted in coverage of the whole sequence except for a short fragment T3 (3 amino acids), which was not detected. It also allowed the insertion of a Pro residue in position 59 to be identified. The results obtained provide evidence for the lack of a substantial level of glycosylation or other post‐translational modifications of subunit 1Dx2, and demonstrate that MALDI‐MS is the most useful method presently available for the direct verification of the gene‐derived sequences of HMW glutenin subunits and similar proteins. Copyright © 2002 John Wiley & Sons, Ltd.