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Istituto di scienza dell'alimentazione

Torna all'elenco Contributi in rivista anno 2005

Contributo in rivista

Tipo: Articolo in rivista

Titolo: Proteomics of gluten: mapping of the 1Bx7 glutenin subunit in Chinese Spring cultivar by matrix-assisted laser desorption/ionization

Anno di pubblicazione: 2005

Formato: Cartaceo

Autori: Alberghina G; Cozzolino R; Fisichella S; Garozzo D; Savarino A

Affiliazioni autori: Department of Chemical Sciences, 6 V. le A. Doria, 95125 Catania, Italy CNR Institute for Chemistry and Technology of Polymers, 6 V. le R. Margherita, 95125 Catania, Italy

Autori CNR:

  • ROSARIA COZZOLINO
  • DOMENICO GAROZZO

Lingua: inglese

Abstract: The verification of the cDNA-deduced sequence of the high molecular weight glutenin subunit 1Bx7 in Chinese Spring cultivar was achieved by direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) analysis of the tryptic fragments. The published sequence of the 1Bx7 subunit contains 5 Lys and 15 Arg residues but, due to the presence of three Arg-Pro bonds, which are generally resistant to cleavage by trypsin, or cleaved to a very limited extent by trypsin, 19 peptides can be predicted. The identification of the tryptic fragments was achieved by direct MALDI-MS analysis by using three different matrices (DHB, SA and HCCA) in combination with the most compatible sample preparation procedures in order to obtain the maximum sequence coverage. MALDI analysis of the 1Bx7 tryptic digest resulted in the identification of the expected peptides and additional fragments arising from non-specific cleavages; the fragments that were not detected are peptides with low mass (from 147.2 to 317.4), so we obtained a sequence coverage of 98.8%. The results reported here also indicated that the sequence of the 1Bx7 subunit from cv. Chinese Spring is different from the cDNA-deduced sequence reported in the literature; in particular, a possible insertion of the hexapeptide QPGQGQ within the sequence Gln630-Tyr725 was suggested. Finally, it is possible to rule out glycosylation of the 1Bx7 subunit, or any other post-translational modification, to within the detection limits of the method.

Lingua altro abstract: inglese

Pagine da: 2069

Pagine a: 2074

Rivista:

RCM. Rapid communications in mass spectrometry Heyden,
Paese di pubblicazione: Regno Unito
Lingua: inglese
ISSN: 0951-4198

Numero volume: 19

Numero fascicolo: 14

DOI: 10.1002/rcm.2032

Indicizzato da: ISI Web of Science (WOS) [000230666700022]

Strutture CNR:

Moduli:

Allegati: Proteomics of gluten: mapping of the 1Bx7 glutenin subunit in Chinese Spring cultivar by matrix-assisted laser desorption/ionization (application/pdf)

 
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