Casein phosphoproteome: Identification of phosphoproteins by combined mass spectrometry and two‐dimensional gel electrophoresis

Mamone, Gianfranco; Caira, Simonetta; Garro, Giuseppina; Nicolai, Adalgisa; Ferranti, Pasquale; Picariello, Gianluca; Malorni, Antonio; Chianese, Lina; Addeo, Francesco

doi:10.1002/elps.200305545

Cited by 55 publications

(42 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Effects of lipid-deprived diet on the O-glycosylation of secretory proteins The sugar motifs covalently bound to k-cas were defined in the early 1980s but the composition of the O-glycans in bovine and sheep k-cas has recently been re-visited using mass spectrometry (Mamone et al, 2003;Holland et al, 2005). The saccharides found in the antennae were essentially galactose, Lipid restriction perturbs protein O-glycosylation N-acetylgalactosamine and N-acetylneuraminic acid (in bovine: NeuAca(2-3)Galb(1-3)[NeuAca(2-6)]GalNAc).…”

Section: Discussionmentioning

confidence: 99%

Lipid-deprived diet perturbs O-glycosylation of secretory proteins in rat mammary epithelial cells

Lavialle¹,

Chanat²

2008

Animal

View full text Add to dashboard Cite

Nutrition modulates both production and composition of milk. Milk composition was studied in rats chronically fed a diet without additional lipids, and therefore eating only traces of the recommended supply of essential polyunsaturated fatty acid. Despite a large decrease in milk-protein synthesis, only protein composition, but not protein concentration, was found to change in the milk of rats following a lipid-deprived diet. Correlatively, we observed a substantial increase in the lactose concentration of milk. Analysis of milk proteins by two-dimensional electrophoresis demonstrated that the relative proportion of the various molecular forms of k-casein, an O-glycosylated protein, was modified in the milk of rats receiving the lipiddeprived diet. In tissues, differences in the two-dimensional pattern of k-casein between control and lipid-deprived rats were similar, if not identical. In contrast to k-casein, the molecular forms of a-lactalbumin, an N-glycosylated protein, were not affected by the diet. These data provide evidence that O-glycosylation of milk proteins in the secretory pathway of mammary epithelial cells is modulated by the lipid content of experimental diets.

show abstract

Section: Discussionmentioning

confidence: 99%

Lipid-deprived diet perturbs O-glycosylation of secretory proteins in rat mammary epithelial cells

Lavialle¹,

Chanat²

2008

Animal

View full text Add to dashboard Cite

show abstract

“…[19] The casein fraction was dissolved in 0.4% NH 4 HCO 3 aqueous solution, pH 8.5, at a concentration of 1 µg/µl and a 20 U enzyme/mg protein ratio was used. The solution was incubated at 37…”

Section: Preparation Of Milk and Enzymatic Dephosphorylation Of Caseimentioning

confidence: 99%

Sequence determination of α_s1‐casein isoforms from donkey by mass spectrometric methods

Cunsolo¹,

Cairone²,

Fontanini³

et al. 2009

J. Mass Spectrom.

View full text Add to dashboard Cite

Four co-eluting components, with experimentally measured M(r) of 23 658, 23 786, 24 278 and 24 406 Da, were detected by reversed-phase high-performance liquid chromatography (RP-HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis in the dephosphorylated casein fraction of a milk sample collected at middle lactation stage from an individual donkey belonging to the Ragusano breed. By coupling RP-HPLC, two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), enzymatic digestions, MALDI-TOF MS and capillary RP-HPLC/nano-electrospray ionization tandem mass spectrometry (nESI-MS/MS) analyses, the four components were identified as donkey's alpha(s1)-CNs and their sequences completely characterized, using the known mare's alpha(s1)-CN (GenBank Acc. No. AAK83668; M(r) 23750.7 Da) as reference. The proteins with M(r) of 23 786 and 23 658 Da differ in the presence of a glutamine residue at position 83 in the full-length component and present the amino acid substitutions Q(8)-->H and H(115)-->Y with respect to the mare's alpha(s1)-CN. The other two components with M(r) 24 406 and 24 278 Da, which also differ in the presence of a glutamine residue at position 88 in the full-length component, show the insertion of the pentapeptide HTPRE between Leu(33) and the Glu(34). The two alpha(s1)-CNs bearing the pentapeptide insertion were named variants A (202 amino acids; M(r) 24 406) and A(1) (201 amino acids; M(r) 24 278), whereas the two alpha(s1)-CNs without the pentapeptide were named variants B (197 amino acids; M(r) 23 786) and B(1) (196 amino acids; M(r) 23 658).

show abstract

“…12 Several studies have shown that in goat milk each a s2 -Cn variant can exist in two forms, identical to those described in ovine milk. For each variant, the main component corresponds to the full length of a s2 -Cn, while the minor component, originated from a skipping event, lacks the internal sequence Glu 34 -Glu 42 or Lys 35 -Glu 43 . 9 The shorter forms of a s2 -Cn variants, as those of a s1 -Cn discussed above, are named here sub-variants.…”

mentioning

confidence: 99%

Detection and characterization by high‐performance liquid chromatography and mass spectrometry of two truncated goat α_s2‐caseins

Cunsolo

Muccilli

Saletti

et al. 2006

Rapid Comm Mass Spectrometry

View full text Add to dashboard Cite

The identification and characterization of truncated forms of goat alphas2-Cn variants A and E are reported. The two proteins, which have experimental Mr values of 24 183 and 24 227 Da, were detected as minor components in a goat milk sample from an autochthonous breed of southern Italy, 'Rossa Mediterranea', by reversed-phase high-performance liquid chromatography/electrospray ionization mass spectrometry (RP-HPLC/ESI-MS). Characterization of the amino acid sequences, performed by coupling trypsin digestion with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), RP-HPLC/ESI-MS and tandem mass spectrometry (MS/MS), demonstrated that the polypeptide chains correspond to the 1-204 sequence of mature alphas2-Cn variant A (component with Mr of 24 183 Da) and E (component with Mr of 24 227 Da), respectively. These components seem to be the product of a differential splicing of pre-messenger RNA during the translation process of the alphas2-Cn variants A and E.

show abstract

Casein phosphoproteome: Identification of phosphoproteins by combined mass spectrometry and two‐dimensional gel electrophoresis

Cited by 55 publications

References 36 publications

Lipid-deprived diet perturbs O-glycosylation of secretory proteins in rat mammary epithelial cells

Lipid-deprived diet perturbs O-glycosylation of secretory proteins in rat mammary epithelial cells

Sequence determination of α_s1‐casein isoforms from donkey by mass spectrometric methods

Detection and characterization by high‐performance liquid chromatography and mass spectrometry of two truncated goat α_s2‐caseins

Contact Info

Product

Resources

About

Casein phosphoproteome: Identification of phosphoproteins by combined mass spectrometry and two‐dimensional gel electrophoresis

Cited by 55 publications

References 36 publications

Lipid-deprived diet perturbs O-glycosylation of secretory proteins in rat mammary epithelial cells

Lipid-deprived diet perturbs O-glycosylation of secretory proteins in rat mammary epithelial cells

Sequence determination of αs1‐casein isoforms from donkey by mass spectrometric methods

Detection and characterization by high‐performance liquid chromatography and mass spectrometry of two truncated goat αs2‐caseins

Contact Info

Product

Resources

About

Sequence determination of α_s1‐casein isoforms from donkey by mass spectrometric methods

Detection and characterization by high‐performance liquid chromatography and mass spectrometry of two truncated goat α_s2‐caseins