Isolation and properties of a new anticoagulant protein from commercial bovine testicular hyaluronidase.
Bovine testicular hyaluronidase from various commercial sources showed the presence of an inhibitor of human plasma prothrombin time (PT). A testicular anticoagulant protein (TAP) was isolated from it by a 3-step procedure. The material was first passed through conconavalin-A-sepharose affinity chromatography where the anticoagulant material was separated from the hyaluronidase  and protease which were retained by the column. In the second step the lower molecular weight proteins were removed by ultrafiltration. The supernatant which contained the anticoagulant protein was passed through the carboxymethyl cellulose column and the active material was eluted by 0.4 M NaCl solution. Sodium dodecyl sulfate (SDS) gel electrophoresis gave a molecular weight of approximately 35000. Unlike many small molecular weight proteins from bovine testes, TAP looses its anticoagulant property by heating for 30 minutes at 55 degrees C or by storage at pH 3.0 for 2 hours and it does not inhibit trypsin or thrombin. Its isoelectric pH was 9.7. Diisopropyl fluorophosphate (DFP) treated TAP was not effective as an anticoagulant.
D000818:Animals; D000925:Anticoagulants / Q000302:isolation & purification*; D002417:Cattle; D004791:Enzyme Inhibitors; D006821:Hyaluronoglucosaminidase / Q000032:analysis*; D008297:Male; D011506:Proteins / Q000302:isolation & purification*; D013735:Testicular Hormones / Q000302:isolation & purification* / Q000494:pharmacology; D013737:Testis / Q000201:enzymology*
D000925:Anticoagulants; D004791:Enzyme Inhibitors; D011506:Proteins; D013735:Testicular Hormones; C039579:testicular anticoagulant protein, Bos taurus; D006821:Hyaluronoglucosaminidase