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Istituto di scienza dell'alimentazione

Torna all'elenco Contributi in rivista anno 2001

Contributo in rivista

Tipo: Articolo in rivista

Titolo: Mass spectrometry-based procedure for the identification of ovine casein heterogeneity

Anno di pubblicazione: 2001

Formato: Cartaceo

Autori: Ferranti Pasquale, Pizzano Rosa, Garro Giuseppina, Caira Simonetta, Chianese Lina, Addeo Francesco

Affiliazioni autori: Pizzano Rosa, Simonetta Caira: Istituto di scienza dell'alimentazione Ferranti Pasquale, Garro Giuseppina, Lina Chianese, Addeo Francesco: Dipartimento di Scienza degli Alimenti - Università degli Studi di Napoli Federico II

Autori CNR:

  • SIMONETTA CAIRA
  • ROSA PIZZANO

Lingua: inglese

Abstract: The efficiency of reversed-phase HPLC, capillary electrophoresis (CE), PAGE and isoelectric focusing with immunoblotting in separating ovine caseins has been evaluated. The assessment was carried out by employing electrospray ionization-mass spectrometry (ESI-MS) and matrix-assisted laser desorption ionization-time of flight as reference tools for identifying protein components. Ovine casein was fractionated by HTPC into four major peaks. With ESI-MS, each peak contained components belonging to only one of the four casein families. On-line liquid chromatography-ESI-MS allowed us to determine each fraction's composition by detecting thirteen alphas1-, eleven alphas2-, seven beta-, and three kappa-casein (CN) components. The alphas1-CN and alphas2-CN consisted of eight and two protein chains respectively of lengths differing through the deletion of one or more peptide sequences; they were also discretely phosphorylated as kappa-CN and beta-CN. By CE at pH 2.5, each casein fraction was as heterogeneous as that resulting from ESI-MS for the single HPLC-derived fractions. The separation of alphas1-CN and alphas2-CN proved to be excellent, with the exception of a co-migration of kappa0-CN with a minor alphas1-CN component and of a glycosylated kappa-CN for with low-phosphorylated alphas1-CN and beta-CN components. Dephosphorylation of whole casein was used to reduce the heterogeneity of the native fractions and by applying currently used analytical techniques it was possible to visualize the protein moiety difference along the CE profile. CE, HPLC, and immunoblotting were all equally capable of effecting an accurate separation of the four dephosphorylated casein families. The spectra obtained by ESI-MS directly on dephosphorylated whole ovine casein samples contained the signals of the four casein families and the relative alphas1-CN variants, the non-allelic alphas1-CN and alphas2-CN forms, dimeric kappa-CN and other newly formed peptides. We suggest using this procedure for rapid characterization of whole casein.

Lingua abstract: inglese

Pagine da: 35

Pagine a: 51

Pagine totali: 17

Rivista:

Journal of dairy research Empire Marketing Board, Dairy Research Committee,
Paese di pubblicazione: Regno Unito
Lingua: inglese
ISSN: 0022-0299

Numero volume: 68

Numero fascicolo: 1

DOI: 10.1017/S0022029900004611

Referee: Sì: Internazionale

Indicizzato da:

  • ISI Web of Science (WOS) [000167519600005]
  • Scopus [2-s2.0-0035096881]
  • PubMed [11289268]

Parole chiave:

  • ovine casein
  • casein heterogeneity
  • electrospray ionization-mass spectrometry
  • capillary electrophoresis

Strutture CNR:

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