|Home | English version | Mappa | Commenti | Sondaggio | Staff | Contattaci||Cerca nel sito|
|Istituto di scienza dell'alimentazione|
Contributo in rivista
Tipo: Articolo in rivista
Titolo: Characterization of wheat gliadin proteins by combined two-dimensional gel electrophoresis and tandem mass spectrometry.
Anno di pubblicazione: 2005
Formato: Elettronico Cartaceo
Autori: Mamone G, Addeo F, Chianese L, Di Luccia A, De Martino A, Nappo A, Formisano A, De Vivo P, Ferranti P.
Affiliazioni autori: Istituto di Scienze dell'Alimentazione del CNR, Avellino, Italy Dipartimento di Scienza degli Alimenti, Università degli Studi di Napoli "Federico II", Portici, Italy Dipartimento di Produzione Animale, Università degli Studi Bari, Bari, Italy
Abstract: A proteomics-based approach was used for characterizing wheat gliadins from an Italian common wheat (Triticum aestivum) cultivar. A two-dimensional gel electrophoresis (2-DE) map of roughly 40 spots was obtained by submitting the 70% alcohol-soluble crude protein extract to isoelectric focusing on immobilized pH gradient strips across two pH gradient ranges, i.e., 3-10 or pH 6-11, and to sodium dodecyl sulfate-polyacrylamide electrophoresis in the second dimension. The chymotryptic digest of each spot was characterized by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and nano electrospray ionization-tandem mass spectrometry (MS/MS) analysis, providing a "peptide map" for each digest. The measured masses were subsequently sought in databases for sequences. For accurate identification of the parent protein, it was necessary to determine de novo sequences by MS/MS experiments on the peptides. By partial mass fingerprinting, we identified protein molecules such as alpha/beta-, gamma-, omega-gliadin, and high molecular weight-glutenin. The single spots along the 2-DE map were discriminated on the basis of their amino acid sequence traits. alpha-Gliadin, the most represented wheat protein in databases, was highly conserved as the relative N-terminal sequence of the components from the 2-DE map contained only a few silent amino acid substitutions. The other closely related gliadins were identified by sequencing internal peptide chains. The results gave insight into the complex nature of gliadin heterogeneity. This approach has provided us with sound reference data for differentiating gliadins amongst wheat varieties.
Lingua altro abstract: inglese
Pagine da: 2859
Pagine a: 2865
Pagine totali: 7
Numero volume: 5(11)
Referee: Sì: Internazionale
Indicizzato da: ISI Web of Science (WOS) 
|Home | Il CNR | I servizi | News | Eventi | Istituti | Focus|