Chemical and immunochemical characterization of caseins and the major whey proteins of rabbit milk.
Caseins were separated from whey proteins by acid precipitation of skimmed rabbit milk. Whole casein was resolved by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis into three major bands with apparent relative molecular masses (Mr of 31 000, 29 000 and 25 000. On agarose/urea-gel electrophoresis whole casein gave three bands with electrophoretic mobilities alpha, beta and gamma. The three components were purified by DEAE-cellulose chromatography under denaturing and reducing conditions. Each was shown to have a different amino acid, hexose and phosphorus content, as well as non-identical peptide fragments after proteinase digestion. The 31 000 Da (dalton) protein, of alpha-electrophoretic mobility, had a high phosphorus content (4.38%, w/w); the 29 000 Da peptide, of gamma-mobility, had the highest hexose content (2.2%, w/w), contained 0.8 cysteine residue per 100 amino acid residues and was susceptible to chymosin digestion corresponding thus to kappa-casein; the 25 000 Da protein migrated to the beta-position. The rabbit casein complex is composed of at least three caseins, two of which (alpha- and kappa-caseins) are analogous to the caseins from ruminants. Although caseins are poor immunogens, specific antibodies were raised against total and purified polypeptides. The antiserum directed against whole casein recognized each polypeptide, each casein corresponding to a distinct precipitation line. The antisera directed against each casein polypeptide reacted exclusively with the corresponding casein and no antiserum cross-reaction occurred between the three polypeptides. From whey, several proteins were isolated, characterized and used as antigens to raise specific antibodies. An iron-binding protein with an apparent Mr of 80 000 was shown to be immunologically and structurally identical with serum transferrin.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002364:Caseins / Q000276:immunology / Q000302:isolation & purification; D055598:Chemical Phenomena; D002621:Chemistry; D004586:Electrophoresis; D000939:Epitopes; D005260:Female; D007122:Immunoelectrophoresis; D008894:Milk Proteins / Q000276:immunology / Q000302:isolation & purification; D010446:Peptide Fragments / Q000032:analysis; D011817:Rabbits
D000596:Amino Acids; D002364:Caseins; D000939:Epitopes; D008894:Milk Proteins; D010446:Peptide Fragments