Two-Dimensional Electrophoresis, MALDI-TOF MS and blotting for identification and characterisation of pathogenesis-related proteins in wheat cultivar

Gy Hajós, E. Szabó, T. Janáky

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3 Citations (Scopus)

Abstract

Protein sets of different wheat cultivars were studied by two-dimensional polyacrylamide gel electrophoresis (2-D-PAGE). One of the wheat cultivars showed several differentially expressed spots, which were submitted to further investigation and identification. The protein spots of interest (several spots of the differentially expressed proteins) were excised from the gel, and digested with trypsin. The digests after purification were analysed by MALDI-TOF MS and the proteins were identified by similarity searching in the protein database. Using this proteomic approach, one chitinase, one inhibitor and one heat shock protein were identified in the water-and salt-soluble extract of a wheat cultivar. PR-protein-related epitopes were found to be allergenic/cross-reactive determinant. This study demonstrated, that the sera of patients suffering from cereal allergy contained IgE antibodies which bound to some polypeptides of the globulin/albumin fraction of wheat flour. Our work describe an array of techniques suitable for characterisation and identification of components of a complex extract of wheat proteins from point of view of food safety and quality and healthy nutrition.

Original languageEnglish
JournalChromatographia
Volume60
Issue numberSUPPL.
Publication statusPublished - 2004

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Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Electrophoresis
Triticum
Proteins
Chitinases
Food Quality
Protein Databases
Allergies
Food Safety
Food safety
Globulins
Electrophoresis, Gel, Two-Dimensional
Flour
Heat-Shock Proteins
Complex Mixtures
Proteomics
Trypsin
Immunoglobulin E
Nutrition
Epitopes

Keywords

  • Blotting
  • Cross-reactive epitopes
  • MALDI-TOF MS
  • Pathogenesis-related proteins
  • Two-dimensional polyacrylamide gel electrophoresis

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry

Cite this

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title = "Two-Dimensional Electrophoresis, MALDI-TOF MS and blotting for identification and characterisation of pathogenesis-related proteins in wheat cultivar",
abstract = "Protein sets of different wheat cultivars were studied by two-dimensional polyacrylamide gel electrophoresis (2-D-PAGE). One of the wheat cultivars showed several differentially expressed spots, which were submitted to further investigation and identification. The protein spots of interest (several spots of the differentially expressed proteins) were excised from the gel, and digested with trypsin. The digests after purification were analysed by MALDI-TOF MS and the proteins were identified by similarity searching in the protein database. Using this proteomic approach, one chitinase, one inhibitor and one heat shock protein were identified in the water-and salt-soluble extract of a wheat cultivar. PR-protein-related epitopes were found to be allergenic/cross-reactive determinant. This study demonstrated, that the sera of patients suffering from cereal allergy contained IgE antibodies which bound to some polypeptides of the globulin/albumin fraction of wheat flour. Our work describe an array of techniques suitable for characterisation and identification of components of a complex extract of wheat proteins from point of view of food safety and quality and healthy nutrition.",
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T1 - Two-Dimensional Electrophoresis, MALDI-TOF MS and blotting for identification and characterisation of pathogenesis-related proteins in wheat cultivar

AU - Hajós, Gy

AU - Szabó, E.

AU - Janáky, T.

PY - 2004

Y1 - 2004

N2 - Protein sets of different wheat cultivars were studied by two-dimensional polyacrylamide gel electrophoresis (2-D-PAGE). One of the wheat cultivars showed several differentially expressed spots, which were submitted to further investigation and identification. The protein spots of interest (several spots of the differentially expressed proteins) were excised from the gel, and digested with trypsin. The digests after purification were analysed by MALDI-TOF MS and the proteins were identified by similarity searching in the protein database. Using this proteomic approach, one chitinase, one inhibitor and one heat shock protein were identified in the water-and salt-soluble extract of a wheat cultivar. PR-protein-related epitopes were found to be allergenic/cross-reactive determinant. This study demonstrated, that the sera of patients suffering from cereal allergy contained IgE antibodies which bound to some polypeptides of the globulin/albumin fraction of wheat flour. Our work describe an array of techniques suitable for characterisation and identification of components of a complex extract of wheat proteins from point of view of food safety and quality and healthy nutrition.

AB - Protein sets of different wheat cultivars were studied by two-dimensional polyacrylamide gel electrophoresis (2-D-PAGE). One of the wheat cultivars showed several differentially expressed spots, which were submitted to further investigation and identification. The protein spots of interest (several spots of the differentially expressed proteins) were excised from the gel, and digested with trypsin. The digests after purification were analysed by MALDI-TOF MS and the proteins were identified by similarity searching in the protein database. Using this proteomic approach, one chitinase, one inhibitor and one heat shock protein were identified in the water-and salt-soluble extract of a wheat cultivar. PR-protein-related epitopes were found to be allergenic/cross-reactive determinant. This study demonstrated, that the sera of patients suffering from cereal allergy contained IgE antibodies which bound to some polypeptides of the globulin/albumin fraction of wheat flour. Our work describe an array of techniques suitable for characterisation and identification of components of a complex extract of wheat proteins from point of view of food safety and quality and healthy nutrition.

KW - Blotting

KW - Cross-reactive epitopes

KW - MALDI-TOF MS

KW - Pathogenesis-related proteins

KW - Two-dimensional polyacrylamide gel electrophoresis

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