Binding of bovine brain tissue factor to concanavalin A-Sepharose. Partial purification of coagulant, arylamidase, and alkaline phosphatase activities.
Tissue factor coagulant activity is adsorbed onto concanavalin A-Sepharose from sodium deoxycholate extracts of delipidated bovine brain powders. Coagulant activity is eluted with alpha-methyl-D-glucoside in sodium deoxycholate with 2--25-fold purification. This material has the same coagulant specific activity as that previously prepared in this laboratory. Alkaline phosphatase and alanyl-beta-naphthylamidase activities in the detergent extract also bind to concanavalin A-Sepharose and elute under the same conditions with 4- and 7-fold purification. In addition to these biological activities, the eluate was composed of protein (67.7%), neutral and amino sugars and sialic acid (22.3%), phospholipid (4.5%), uronic acid (3.8%) and nucleic acid (1.7%). This preparation is slightly enriched in carbohydrates compared to previous preparations. Concanavalian A-Sepharose therefore appears to be useful material for partial purification of several mammalian plasma membrane components with retention of biological function.
D000042:Absorption; D000469:Alkaline Phosphatase / Q000378:metabolism*; D000626:Aminopeptidases / Q000378:metabolism*; D000818:Animals; D001059:Apoproteins / Q000378:metabolism; D001921:Brain / Q000201:enzymology* / Q000378:metabolism; D002417:Cattle; D003208:Concanavalin A / Q000378:metabolism*; D003840:Deoxycholic Acid; D066298:In Vitro Techniques; D012685:Sepharose; D013925:Thromboplastin / Q000302:isolation & purification / Q000378:metabolism*
D001059:Apoproteins; D003840:Deoxycholic Acid; D003208:Concanavalin A; D012685:Sepharose; D013925:Thromboplastin; D000469:Alkaline Phosphatase; D000626:Aminopeptidases