Studies on the collagen glucosyltransferase activity present in platelets and plasma.
1. Collagen glucosyltransferase was demonstrated to be associated with pig platelets by using a specific assay for the synthesis of [(14)C]glucosylgalactosylhydroxylysine. 2. This enzyme from pig platelets required denatured collagen as substrate and the reaction was not inhibited by the presence of triple-helical collagen. These observations indicate that the platelet enzyme cannot form either an enzyme-substrate complex or an enzyme-inhibitor complex with triple-helical collagen. 3. Platelets were fractionated by sucrose-density-gradient centrifugation after either lysis by a glycerol-loading technique or homogenization. Assays of subcellular fractions for collagen glucosyltransferase activity indicated that the enzyme was localized predominantly in the cytosolic fraction and less than 5% of the activity was associated with the membrane fractions. 4. Enzyme assays were carried out on platelet-rich plasma and platelet-poor plasma prepared from pig and human blood. These analyses indicated that most of the collagen glucosyltransferase activity of platelet-rich plasma was in a soluble form and only about 10% was associated with platelets. 5. Comparative studies on the enzyme activity in plasma and platelets of various animal species revealed marked variation, with the guinea pig exhibiting the highest activity. In most cases there was a correlation between the activity found in platelets and plasma, but little species variation was noted in enzyme amounts detected in bone-marrow preparations. 6. The results described here are discussed in the context of the proposal that collagen glucosyltransferase might play a role in mediating collagen-platelet adhesion.
D000818:Animals; D001792:Blood Platelets / Q000201:enzymology*; D001853:Bone Marrow / Q000201:enzymology; D002417:Cattle; D003094:Collagen; D004285:Dogs; D005964:Glucosyltransferases / Q000097:blood*; D006168:Guinea Pigs; D006801:Humans; D051379:Mice; D011487:Protein Conformation; D011817:Rabbits; D051381:Rats; D012756:Sheep; D013045:Species Specificity; D013347:Subcellular Fractions / Q000201:enzymology; D013552:Swine
D003094:Collagen; D005964:Glucosyltransferases