Purification and characterization of a differentiation-specific sialoglycoprotein of lactating-guinea-pig mammary tissue.
A large acidic glycoprotein, PAS-I, was purified from the fat-globule membrane of guinea-pig milk. Threonine and serine accounted for over 30 mol% of the amino acids, and galactose, N-acetylgalactosamine, N-acetylglucosamine, mannose and sialic acid were the principal sugars detected. On a molar basis, sialic acid accounted for over 60% of the total sugar. Removal of sialic acid by treatment with neuraminidase revealed the presence of binding sites for peanut (Arachis hypogaea) agglutinin, a lectin specific for the sugar sequence beta-D-Gal-(beta 1----3)-D-GalNac (the T antigen). The distribution of PAS-I-related epitopes, defined by five monoclonal antibodies, was determined in the mammary gland and in other guinea-pig tissues. PAS-I was maximally expressed on the apical surfaces of secretory cells in lactating mammary tissue and was either absent, or present in much lower amounts, in the glands of virgin or pregnant animals. PAS-I epitopes were not detected in liver, heart, spleen, pancreas, ovary, uterus, lung or intestine, either by immunofluorescence microscopy or by immunoblotting techniques. Several of the PAS-I-specific antibodies bound to mucins of high Mr in human fat-globule membrane, and similarities and differences between PAS-I and the human mucins are discussed. PAS-I and epitopes of this glycoprotein will be useful as indicators of differentiation in mammary cells and of markers of the apical surface of these cells during lactation.
Johnson
V G
VG
Department of Animal Sciences, University of Maryland, College Park 20742.
Greenwalt
D E
DE
Madara
P J
PJ
Mather
I H
IH
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D002241:Carbohydrates / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D005260:Female; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D006801:Humans; D007774:Lactation / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000032:analysis; D008562:Membrane Glycoproteins / Q000276:immunology / Q000302:isolation & purification*; D008566:Membranes / Q000032:analysis; D008894:Milk Proteins / Q000302:isolation & purification*; D011247:Pregnancy; D014018:Tissue Distribution
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D000939:Epitopes; D008562:Membrane Glycoproteins; D008894:Milk Proteins