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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: 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

Autori CNR:

  • GIANFRANCO MAMONE

Lingua: inglese

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

Rivista:

Proteomics Wiley-VCH-Verl.
Paese di pubblicazione: Germania
Lingua: inglese
ISSN: 1615-9853

Numero volume: 5(11)

DOI: 10.1002/pmic.200401168

Referee: Sì: Internazionale

Indicizzato da: ISI Web of Science (WOS) [000231036100015]

Parole chiave:

  • Gliadin
  • Peptide mass fingerprinting
  • Tandem mass spectrometry
  • Wheat

URL: http://www.isa.cnr.it

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