Cereal n-glycoproteins Enrichment by Lectin Affinity Monolithic Chromatography
Research of cereal glycoproteins is important for understanding of their functional properties, their role during technological processing of cereals and it can serve as a valuable tool for the detection of cereal allergens. The main intention of this study was the screening of profile of water-soluble glycoproteins present in barley (Hordeum vulgare), wheat durum (Triticum durum) and spelt (Triticum spelta). Lectin monolithic HPLC column was used for rapid and effective enrichment of glycoprotein fractions. Captured glycoproteins were electrophoretically separated and analyzed by MALDI-TOF MS. Presented procedure resulted in identification of a group of N-glycoprotein candidates with affinity to lectin concanavalin A (ConA). Such molecules could have, among others, an allergenic potential. Majority of captured low-molecular-weight glycoproteins belong to alpha amylase/trypsin inhibitors family. However, most of the higher-molecular-weight proteins identified in lectin bound fractions have not been described as glycoproteins yet. Obtained results improved the knowledge about (glyco)protein content in cereal grain. The connection of lectin HPLC-GE-MS was proved as a convenient strategy for identification of cereal glycoproteins. Suggested method is universal and can be applied for various cereals and food-stuffs.
Keywordsbarley wheat glycoprotein mass spectrometry lectin chromatography
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This work was supported by the Grant Agency of the Czech Republic (Grant No. P503/12/P395) and by the Institutional Research Plan RVO:68081715 of the Institute of Analytical Chemistry of the CAS, v. v. i., Czech Republic.
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