title: Highly phosphorylated region of chicken riboflavin-binding protein: chemical characterization and 31P NMR studies.
abstract: Phosphate residues on chicken riboflavin-binding protein (RBP) have been implicated in the recognition and deposition of the protein-vitamin complex in egg yolk by the ovary. We demonstrate that all of the phosphate of RBP is linked to serine and that most if not all of the phosphoserine residues are contained in a single tryptic peptide having a composition SerP8, Glx6, His2, Leu2, Ala, Met, Lys. Despite differences in the tissue of synthesis, RBPs isolated from egg yolk and egg white yield phosphopeptides virtually identical in their amino acid composition and 101-MHz 31P NMR spectra. This implies that posttranslational phosphorylation of RBP is the same in liver and oviduct. The 31P NMR spectrum of the phosphopeptide is quite different from that of the phosphoprotein. The 31P NMR spectra of egg yolk and egg white RBP are quite similar but not identical and are unaffected by the binding of riboflavin. Optimal resolution of seven phosphorus resonances was obtained near pH 7.0. The titration behavior of all the phosphoserine residues is similar. We propose that this highly anionic peptide, exposed at the surface of RBP, is the principal determinant for the uptake of RBP by the vitelline membrane of the ovarian follicle.
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D004529:Egg White; D004530:Egg Yolk; D005260:Female; D009682:Magnetic Resonance Spectroscopy; D026901:Membrane Transport Proteins; D009865:Oocytes / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis; D010748:Phosphopeptides / Q000302:isolation & purification; D012256:Riboflavin / Q000378:metabolism; D014357:Trypsin
chemical_list: D000596:Amino Acids; D002352:Carrier Proteins; D026901:Membrane Transport Proteins; D010446:Peptide Fragments; D010748:Phosphopeptides; C019079:riboflavin-binding protein; D014357:Trypsin; D012256:Riboflavin