Isolation and characterization of proteoglycans from bovine lung.
Proteoglycans were extracted from bovine lung gas exchange tissue, pleura, and bronchioles with 4.0 M guanidinium chloride at 5 degrees C in the presence of protease inhibitors. Preliminary purification of the proteoglycans was achieved by an initial CsCl isopycnic centrifugation (rho 0 = 1.33) and through precipitation with cetylpyridinium chloride in 0.5 M KCl. Further purification and fractionation of proteoglycans was achieved by a second CsCl isopycnic centrifugation (rho 0 = 1.45) in 4.0 M guanidinium chloride. Based on the ultracentrifuge profiles and electrophoretic behavior, the major fractions were pooled. They were purified further by gel filtration on Sepharose CL-2B and characterized by extensive analyses. A heparan sulfate proteoglycan was the major proteoglycan identified in the gas exchange tissue and in the pleura. The major proteoglycan component from the bronchioles was a chondroitin sulfate proteoglycan. Approximate molecular weight of 2 x 10(6) for the heparan sulfate proteoglycan from the pleura and chondroitin sulfate proteoglycan from the bronchioles and 1 x 10(6) for the heparan sulfate proteoglycan from the gas exchange tissue were estimated from gel filtration analyses. After incubation with hyaluronic acid, the chondroitin sulfate proteoglycans from the bronchioles showed an increase in specific viscosity and a higher molecular weight compound eluting near the void volume in Sepharose CL-2B column chromatography. The proteoglycans exhibited varied anticoagulant activities in Stypven, partial thromboplastin and thrombin clotting times and inhibited thrombin-induced platelet aggregation.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D002417:Cattle; D006820:Hyaluronic Acid / Q000378:metabolism; D008168:Lung / Q000032:analysis*; D008722:Methods; D010974:Platelet Aggregation / Q000187:drug effects; D011509:Proteoglycans / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D013917:Thrombin / Q000037:antagonists & inhibitors
D000596:Amino Acids; D011509:Proteoglycans; D006820:Hyaluronic Acid; D013917:Thrombin