title: Cartilage fucoproteins with sites for cross-linking by transglutaminase.
abstract: Slices of various types of cartilage were incubated with either L-[6-3H]fucose or [1,4-3H(N)]putrescine. Homogenization of the slices and fractionation of the homogenates showed for both labels that an insoluble collagenase-resistant fraction had the highest specific activity (dpm/mg dry weight). Examination of an exhaustive proteolytic digest of this insoluble fraction by ion-exchange high performance liquid chromatography showed the presence of gamma-glutamyl[3H]putrescine. Chromatography of solubilized [3H]fucoprotein fractions showed the presence of several low molecular weight peaks, as well as high molecular weight material. Incubation of [3H]fucoprotein extracts with transglutaminase increased the high molecular weight peaks and decreased the low molecular weight ones. Incubation of the cartilage slices with L-[3H]fucose plus 0.5 mM dansylcadaverine, an inhibitor of transglutaminase, caused a decrease in the insoluble and high molecular weight fraction relative to the low molecular weight peaks. It is hypothesized that this is due to inhibition of cross-link formation between fucoprotein components of the cartilage which are transglutaminase substrates. One major low molecular weight peak, which labels with both fucose and putrescine, corresponds in size with the 15,000 subunit of collagen III aminopropeptide, which is known to be a substrate for transglutaminase.
mesh_terms: D000818:Animals; D002356:Cartilage / Q000378:metabolism*; D002358:Cartilage, Articular / Q000378:metabolism; D002417:Cattle; D004285:Dogs; D005643:Fucose / Q000378:metabolism; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D011700:Putrescine / Q000378:metabolism; D011817:Rabbits; D011503:Transglutaminases / Q000378:metabolism*; D014316:Tritium
chemical_list: D006023:Glycoproteins; C016465:fucoproteins; D014316:Tritium; D005643:Fucose; D011503:Transglutaminases; D011700:Putrescine