Rapid purification of collagen binding glycoproteins secreted by cultured endothelial cells.
Two distinct lines of bovine aortal endothelial cells and also chick embryo fibroblasts, secrete two major non-associating collagen binding proteins into the growth medium. Both proteins are anionic, mannose-containing glycoproteins but differ in molecular weight (Mr = 220,000 and 70,000 daltons respectively), affinity for heparin and reactivity to antiserum against plasma fibronectin. The large molecular weight glycoprotein was identified as fibronectin. In this paper we present a novel rapid procedure for purification of each glycoprotein under non-denaturing conditions. A simple two step affinity column procedure was used to achieve high yields (80% and 85% respectively) of native, non-denatured glycoproteins suitable for fine analytical purposes and cell interaction studies.
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002642:Chick Embryo; D003094:Collagen / Q000302:isolation & purification* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004727:Endothelium / Q000166:cytology; D005353:Fibronectins / Q000302:isolation & purification; D006023:Glycoproteins / Q000302:isolation & purification*; D008358:Mannose / Q000378:metabolism; D010649:Phenylalanine / Q000378:metabolism; D011392:Proline / Q000378:metabolism; D011489:Protein Denaturation; D011506:Proteins / Q000302:isolation & purification*
C032435:CCA protein, rat; D005353:Fibronectins; D006023:Glycoproteins; D011506:Proteins; D010649:Phenylalanine; D003094:Collagen; D011392:Proline; D008358:Mannose