title: Modulation of two distinct galactosyltransferase activities in populations of mouse peritoneal macrophages.
abstract: We have examined two galactosyltransferase activities in membrane preparations obtained from resident macrophages, from resident macrophages maintained in culture for 24 hr, and from thioglycollate (TG)-elicited macrophages. Transfer of galactose from uridine diphosphate (UDP)-galactose to N-acetylglucosamine is 2.6 times higher in membranes prepared from TG macrophages (107 +/- 5.5 nmol/hr/mg) than in membranes prepared from resident macrophages (41 +/- 2.0 nmol/hr/mg). Membranes obtained from resident macrophages cultured for 24 hr exhibit a 2.5 times higher activity (102 +/- 4.4 nmol/hr/mg) than membranes from resident cells plated for 4 hr. Transferase activity in membranes derived from TG macrophages is not significantly affected by overnight culture. The transferase reaction product, isolated on Bio-Gel P-4 and analyzed by galactosidase treatments, was identified as galactosyl-beta 1, 4-N-acetylglucosamine. The enzyme, therefore, is UDP-galactose:2-acetamido-2-deoxy-D-glucose 4 beta-galactosyltransferase. This is supported by the fact that this galactosyltransferase activity is specifically inhibited by high concentrations of N-acetylglucosamine (200 mM). We have also examined the transfer of galactose to N-acetyllactosamine. Membranes from TG-elicited macrophages contain a UDP-galactose:galactosyl-beta 1, 4-N-acetylglucosamine 3 alpha-galactosyltransferase which synthesizes the trisaccharide, galactosyl-alpha 1, 3-galactosyl-beta 1,4-N-acetylglucosamine. This product was identified by gel filtration chromatography, high performance liquid chromatography, and galactosidase digestions. This alpha-galactosyltransferase activity was not detected in membranes prepared from resident macrophages. These results indicate that glycosyltransferase activities are modulated in populations of mouse macrophages, and that these changes correlate with changes in cell surface lactosaminoglycans reported previously.
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D005690:Galactose / Q000378:metabolism; D005700:Galactosyltransferases / Q000378:metabolism*; D007788:Lactose Synthase / Q000378:metabolism; D008264:Macrophages / Q000201:enzymology*; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL / Q000378:metabolism; D010529:Peritoneal Cavity / Q000166:cytology; D013864:Thioglycolates / Q000494:pharmacology; D006022:beta-N-Acetylglucosaminylglycopeptide beta-1,4-Galactosyltransferase / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D013864:Thioglycolates; C017487:2-mercaptoacetate; D005700:Galactosyltransferases; D007788:Lactose Synthase; D006022:beta-N-Acetylglucosaminylglycopeptide beta-1,4-Galactosyltransferase; D000117:Acetylglucosamine; D005690:Galactose