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Primary structure of ovine αsl-caseins: localization of phosphorylation sites and characterization of genetic variants A, C and D*

Published online by Cambridge University Press:  01 June 2009

Pasquale Ferranti ,
Antonio Malorni ,
Gianpaolo Nitti ,
Pasquale Laezza ,
Rosa Pizzano ,
Lina Chianese  and
Francesco Addeo
Pasquale Ferranti
Affiliation:
Dipartimento di Scienza degli Alimenti,, Facoltà di AgrariaUniversità degli Studi di Napoli Federico II, Parco Gussone, 80055 Portici, Italia
Antonio Malorni
Affiliation:
Servizio di Spettrometria di Massa, CNR, Via Pansini 5, 80131 Napoli, Italia
Gianpaolo Nitti
Affiliation:
CEINGE, Via Pansini 5, 80131 Napoli, Italia
Pasquale Laezza
Affiliation:
Dipartimento di Scienza degli Alimenti,, Facoltà di AgrariaUniversità degli Studi di Napoli Federico II, Parco Gussone, 80055 Portici, Italia
Rosa Pizzano
Affiliation:
Dipartimento di Scienza degli Alimenti,, Facoltà di AgrariaUniversità degli Studi di Napoli Federico II, Parco Gussone, 80055 Portici, Italia
Lina Chianese
Affiliation:
Dipartimento di Scienza degli Alimenti,, Facoltà di AgrariaUniversità degli Studi di Napoli Federico II, Parco Gussone, 80055 Portici, Italia
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Summary

The primary structures of ovine α>s1-casein variants A, C and D (formerly called Welsh variant) were determined. Separation of variants from whole casein was achieved using a fast and reliable reversed-phase HPLC method. Extended structural characterization of the purified proteins using electrospray mass spectrometry, automated Edman degradation and peptide mapping by means of HPLC-fast atom bombardment-mass spectrometry demonstrated that the mature protein was a mixture of two molecular species that differed in the deletion of residues 141–148 and were therefore 199 and 191 residues long respectively. The 199 residue peptide chain, which accounted for ∼ 80% of the entire translated αsl-casein, was as long as its caprine and bovine counterparts, and had a 98 and 89 % degree of identity with those two proteins respectively. Nine serine residues (positions 12, 44, 46, 64 to 68 and 75) were fully phosphorylated in αsl-casein A, whereas Ser115 and Ser41 were phosphorylated by ∼ 50 and ∼ 20% respectively. The differences between the three genetic variants A, C and D were simple silent substitutions, which however involved the degree to which the protein was phosphorylated. Variant C differed from variant A in the substitution Ser13 →> Pro13 which determined the loss of the phosphate group on site 12 of the protein chain, SerP12→>Ser12. A further substitution, SerP68 →> Asn68 caused the disappearance of both phosphate groups in the phosphorylated residues Ser64 and Ser66 in variant D; in this last casein variant there was no evidence of phosphorylation at Ser41.

Type
Original Articles
Information
Journal of Dairy Research , Volume 62 , Issue 2 , May 1995 , pp. 281 - 296
Copyright
Copyright © Proprietors of Journal of Dairy Research 1995

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