Characterization of phosphate sites in native ovine, caprine, and bovine casein micelles and their caseinomacropeptides: a solid-state phosphorus-31 nuclear magnetic resonance and sequence and mass spectrometric study.
The phosphate sites in native ovine, caprine, and bovine casein micelles have been analyzed using sequence analysis, mass spectrometric analysis, and solid-state 31P nuclear magnetic resonance spectroscopy. Using a combination of S-ethylcysteine derivatization, sequence analysis, and mass spectrometric analysis, the phosphorylation sites of ovine (SerP151 and SerP168), caprine (SerP151 and SerP168), and bovine (SerP149) caseinomacropeptides have been localized. Various solid-state 31P methods using magic angle spinning have been applied to ascertain the local structure and dynamics of the phosphorylated serine residues and the inorganic calcium phosphates within the micelles. Contributions from the phosphorylated serine residues of kappa-CN, located in the C-terminal portion of the molecule, to the mobile constituents of the micelles were assigned by comparison with 31P nuclear magnetic resonance spectra of purified caseinomacropeptides from the various species in the dissolved state. Comparison of the 31P magic angle spinning nuclear magnetic resonance spectra of ovine, caprine, and bovine casein micelles indicates that the micelles from these species are very similar but not identical.
Rasmussen
L K
LK
Protein Chemistry Laboratory, University of Aarhus, Denmark.
Sørensen
E S
ES
Petersen
T E
TE
Nielsen
N C
NC
Thomsen
J K
JK
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis / Q000378:metabolism; D000818:Animals; D002364:Caseins / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle / Q000378:metabolism*; D005260:Female; D006041:Goats / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods / Q000662:veterinary; D008823:Micelles; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D010710:Phosphates / Q000032:analysis*; D010759:Phosphorus Isotopes; D010766:Phosphorylation; D017386:Sequence Homology, Amino Acid; D012756:Sheep / Q000378:metabolism*
D000596:Amino Acids; D002364:Caseins; D008823:Micelles; D010455:Peptides; D010710:Phosphates; D010759:Phosphorus Isotopes