Incorporation of N-fluoroacetyl-D-glucosamine into hyaluronate by rabbit tracheal explants in organ culture.
1. Incubation of rabbit tracheal explants with N-[(3)H]acetyl-d-glucosamine and N-acetyl-d-[1-(14)C]glucosamine led to labelling of a number of soluble macromolecular products separable from the medium, after papain digestion, by ion-exchange chromatography. 2. With N-acetyl-d-[1-(14)C]glucosamine in the incubation medium, a neutral glycoprotein, two acidic glycoprotein fractions, hyaluronic acid and a glycosaminoglycan fraction were obtained and all were radioactively labelled. Similar labelling occurred with N-fluoroacetyl-d-[1-(14)C]glucosamine or N-fluoro[(3)H]acetylglucosamine as precursor. 3. Maximal labelling was obtained at 96h after incubation of cultures. N-Fluoroacetyl-glucosamine under these conditions was incorporated into hyaluronate less efficiently than N-acetylglucosamine. 4. With N-fluoroacetyl-d-[1-(14)C]glucosamine as precursor, a hyaluronate component was separated that on enzymic degradation by glycosidases (hyaluronidase, beta-glucuronidase and N-acetyl-beta-hexosaminidase) yielded a (14)C-labelled oligosaccharide fraction together with N-acetyl-d-[1-(14)C]glucosamine and N-fluoroacetyl-d-[1-(14)C]glucosamine, consistent with some exchange of N-acetyl groups having occurred. 5. The results on enzymic degradation of labelled macromolecules by glycosidases suggest that the presence of incorporated N-fluoroacetyl side chains may render the hyaluronate analogue more resistant to hyaluronidase.
D000117:Acetylglucosamine / Q000031:analogs & derivatives* / Q000378:metabolism; D000818:Animals; D002852:Chromatography, Ion Exchange; D005944:Glucosamine / Q000031:analogs & derivatives*; D006026:Glycoside Hydrolases; D006820:Hyaluronic Acid / Q000096:biosynthesis*; D008297:Male; D009924:Organ Culture Techniques; D011817:Rabbits; D014132:Trachea / Q000378:metabolism*
C022140:N-fluoroacetyl-D-glucosamine; D006820:Hyaluronic Acid; D006026:Glycoside Hydrolases; D005944:Glucosamine; D000117:Acetylglucosamine