Activation of mouse peritoneal macrophages alters the structure and surface expression of protein-bound lactosaminoglycans.
We have begun to analyze and compare the surface carbohydrates present on populations of resident and activated mouse peritoneal macrophages. The activated macrophage populations studied include TG-elicited macrophages, BCG-activated macrophages, and resident macrophages cultured for 24 hr in the presence of lymphokines or heterologous serum. Analysis of glycopeptides generated by pronase digestion of surface glycoproteins labeled by the neuraminidase/galactose oxidase/NaB3H4 method indicates that the macrophage surface contains a class of high m.w. carbohydrates susceptible to degradation by endo-beta-galactosidase, lactosaminoglycans. These lactosaminoglycans are sialylated type 2 carbohydrates containing the repeating lactosamine disaccharide Gal beta 1-4GlcNAc as well as fucose residues. Macrophage activation was observed to markedly alter surface lactosaminoglycans. The alterations observed include 1) an increase in surface expression as determined by both an increase in neuraminidase/galactose oxidase/NaB3H4 labeling and by the ability of activated but not resident macrophages to bind I antibodies as assayed by indirect immunofluorescent surface staining, 2) the addition of alpha-galactose residues, and 3) an increase in GlcNAc beta 1-6Gal branching as indicated by an increased resistance to endo-beta-galactosidase degradation and by the ability of activated macrophages to bind I antibodies. These observations demonstrate that macrophage activation results in specific and substantial alterations in protein-bound surface carbohydrates.
D000606:Amino Sugars / Q000378:metabolism*; D000818:Animals; D002236:Carbohydrate Conformation; D002850:Chromatography, Gel; D005260:Female; D005455:Fluorescent Antibody Technique; D006020:Glycopeptides / Q000378:metabolism; D008262:Macrophage Activation; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009844:Oligosaccharides / Q000378:metabolism; D010529:Peritoneal Cavity / Q000166:cytology; D011134:Polysaccharides / Q000378:metabolism*; D011485:Protein Binding; D001616:beta-Galactosidase / Q000494:pharmacology
D000606:Amino Sugars; D006020:Glycopeptides; D008565:Membrane Proteins; D009844:Oligosaccharides; D011134:Polysaccharides; C039448:lactosaminoglycan; D001616:beta-Galactosidase