title: Delipidation of alpha 1-acid glycoprotein. Propranolol binding to this glycoprotein and its modification by extracted material and exogenous lipids.
abstract: Propranolol binding to human alpha 1-acid glycoprotein (AAG) delipidated by two methods is described. Commercial AAG (99% pure) was either precipitated by ethanol-acetone and then washed by ether, or it was precipitated by ethanol. Binding capacity was quantified by the product n x Ka where n denotes the number of binding sites and Ka the association constant (M-1). Propranolol binding to nondelipidated AAG (n x Ka = 0.113 +/- 0.013 microM-1) was clearly increased after precipitation by ethanol-acetone (n x Ka = 0.386 +/- 0.109 microM-1) or precipitation by ethanol (n x Ka = 0.312 +/- 0.096 microM-1). Binding capacity potentiation cannot be due to modification of AAG microheterogeneity forms, as two-dimensional gel electrophoresis pattern of AAG in presence of concanavalin A was not altered after both methods. Recombination of precipitated AAGs with supernatant dry residue resulted in the abrogation of observed potentiation. Moreover, addition of a polar lipid, linoleic acid, (from 30 to 300 microM) strongly inhibited propranolol binding. These results indicated that glycoprotein precipitation by ethanol provided a simple method to further study binding inhibitors associated with isolated AAG.
authors.affiliation: Department of Pharmacology, Hôpital Cochin, Paris, France.
authors.forename: L
authors.initials: L
authors.lastname: Chauvelot-Moachon
authors.forename: F
authors.initials: F
authors.lastname: Tallet
authors.forename: C
authors.initials: C
authors.lastname: Durlach-Misteli
authors.forename: J P
authors.initials: JP
authors.lastname: Giroud
mesh_terms: D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003208:Concanavalin A / Q000302:isolation & purification; D003956:Dialysis; D004591:Electrophoresis, Polyacrylamide Gel; D000431:Ethanol; D006801:Humans; D007122:Immunoelectrophoresis; D050356:Lipid Metabolism; D009961:Orosomucoid / Q000378:metabolism*; D011433:Propranolol / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D009961:Orosomucoid; D003208:Concanavalin A; D000431:Ethanol; D011433:Propranolol