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Istituto di scienza dell'alimentazione

Torna all'elenco Contributi in rivista anno 2013

Contributo in rivista

Tipo: Articolo in rivista

Titolo: Challenging the heterogeneity of casein by an IEF/MALDI-TOF "virtual 2D-like" approach

Anno di pubblicazione: 2013

Formato: Elettronico Cartaceo

Autori: Mamone, Gianfranco; Caira, Simonetta; Garro, Giuseppina; Mauriello, Rosalba; Nicolai, Maria Adalgisa; Picariello, Gianluca; Calabrese, Maria Grazia; Ferranti, Pasquale; Chianese, Lina; Addeo, Francesco

Affiliazioni autori: Consiglio Nazionale delle Ricerche (CNR); University of Naples Federico II

Autori CNR:

  • SIMONETTA CAIRA
  • PASQUALE FERRANTI
  • GIANFRANCO MAMONE
  • GIANLUCA PICARIELLO

Lingua: inglese

Abstract: A "virtual two-dimensional (2D)-like" approach based on pH 2.5-6.0 gel isoelectrofocusing coupled to MALDI-TOF mass spectrometry (MS) was addressed for challenging the ovine casein heterogeneity due to polymorphism and post-translational modifications. The procedure enabled the identification of the main casein components including i) the four casein families at different degrees of phosphorylation, ii) nonallelic variants of either alpha(s1)- or alpha(s2)-CN, iii) differently glycosylated kappa-CN, and iv) high molecular mass proteolytic peptides. Protein assignment was confirmed by MALDI-TOF (MS) mass mapping of the tryptic digests, while phosphorylation sites were localized by tandem mass spectrometry. A straightforward identification was hindered by unexpected molecular mass shifts due to the oxidization of alpha(s1)- and the beta-CN. Indeed, using nano-ESI-MS, the casein sub-types were proved to contain a variable number of oxidized methionine residues. Data were additionally substantiated using polyclonal antibodies raised against the single casein families for immunolabelling purposes. The outcomes herein demonstrate the potentiality of the "virtual 2D-like" approach and provide indications useful for the inter-laboratory discrepancies in casein identification. They also greatly facilitate the comparison of data and the establishment of multi-user image-based isoelectrofocusing gels. (C) 2012 Elsevier Ltd. All rights reserved.

Lingua abstract: inglese

Pagine da: 1263

Pagine a: 1272

Pagine totali: 10

Rivista:

Food research international Elsevier Applied Science on behalf of the Canadian Institute of Food Science and Technology,
Paese di pubblicazione: Regno Unito
Lingua: inglese
ISSN: 0963-9969

Numero volume: 54

Numero fascicolo: 1

DOI: 10.1016/j.foodres.2012.11.038

Referee: Sė: Internazionale

Stato della pubblicazione: Published version

Indicizzato da: ISI Web of Science (WOS) [000329377700150]

Parole chiave:

  • Virtual 2D-like
  • Casein
  • Isoelectrofocusing
  • MALDI-TOF
  • Phosphorylation

Strutture CNR:

Moduli:

Allegati: Challenging the heterogeneity of casein by an IEF/MALDI-TOF "virtual 2D-like" approach (application/pdf)

 
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