Studies on the structure of milk fat globule membrane.
Milk fat globule membrane was solubilized with sodium dodecyl sulfate and mercaptoethanol and the membrane proteins were separated by SDS-polyacrylamide gel electrophoresis. The membrane preparations contained three major size classes of polypeptide of 155,000, 62,500 and 43,500 daltons. At least five glycopeptides were separated of which two stained intensely with periodic acid-Schiff reagent, but poorly with coomassie blue. Trypsin hydrolysis of whole cream and isolated milk fat globule membrane revealed major differences in the rates of protein hydrolysis. Many of the membrane proteins of whole cream resisted proteolysis compared with the same proteins in the isolated membrane. Two glycopeptides were resistant to trypsin digestion in either preparation. Treatment of whole cream with neuraminidase led to the release of at least 70% of the protein-bound sialic acid. Whole cream and isolated membrane samples were iodinated with 125I in the presence of lactoperoxidase and hydrogen peroxide. The membrane proteins were significantly more accessible to lactoperoxidase-125I i in isolated membrane compared with the proteins of whole cream. Polypeptides of molecular weight 43,500 and approximately 48,000 daltons were predominantly labelled in whole cream and could be eluted from the fat globules with magnesium chloride (1.5m). The results strongly suggest that the proteins of milk fat globule membrane are asymmetrically arranged in the membrane and that most of the protein-bound sialic acid is present on the external surface of milk fat globules.
D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000648:ultrastructure*; D005260:Female; D006020:Glycopeptides / Q000032:analysis; D007784:Lactoperoxidase; D008055:Lipids / Q000032:analysis; D008274:Magnesium; D008321:Mammary Glands, Animal / Q000032:analysis; D008892:Milk / Q000032:analysis; D008894:Milk Proteins / Q000032:analysis; D008970:Molecular Weight; D012794:Sialic Acids / Q000032:analysis; D014357:Trypsin
D006020:Glycopeptides; D008055:Lipids; D008894:Milk Proteins; D012794:Sialic Acids; D007784:Lactoperoxidase; D014357:Trypsin; D008274:Magnesium