title: Tandem purification of two HDL-associated partner proteins in human plasma, paraoxonase (PON1) and phosphate binding protein (HPBP) using hydroxyapatite chromatography.
abstract: Human plasma paraoxonase (PON1) is calcium-dependent enzyme that hydrolyses esters, including organophosphates and lactones, and exhibits anti-atherogenic properties. Human phosphate binding protein (HPBP) was discovered as contaminant during crystallization trials of PON1. This observation and uncertainties for the real activities of PON1 led us to re-evaluate the purity of PON1 preparations. We developed a hydroxyapatite chromatography for the separation of both HDL-associated proteins. We confirmed that: (1) HPBP is strongly associated to PON1 in HDL, and generally both proteins are co-purified; (2) standard purification protocols of PON1 lead to impure enzyme; (3) hydroxyapatite chromatography allows the simultaneous purification of PON1 and HPBP.
authors.affiliation: Unité d'Enzymologie, Département de Toxicologie, Centre de Recherches du Service de Santé des Armées, 38702 La Tronche Cedex, France.
authors.forename: Frédérique
authors.initials: F
authors.lastname: Renault
authors.forename: Eric
authors.initials: E
authors.lastname: Chabrière
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Andrieu
authors.forename: Bernard
authors.initials: B
authors.lastname: Dublet
authors.forename: Patrick
authors.initials: P
authors.lastname: Masson
authors.forename: Daniel
authors.initials: D
authors.lastname: Rochu
mesh_terms: D000595:Amino Acid Sequence; D043303:Aryldialkylphosphatase / Q000097:blood* / Q000737:chemistry; D008076:Cholesterol, HDL / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008969:Molecular Sequence Data; D028044:Phosphate-Binding Proteins / Q000097:blood* / Q000737:chemistry; D012015:Reference Standards
chemical_list: D008076:Cholesterol, HDL; D028044:Phosphate-Binding Proteins; D043303:Aryldialkylphosphatase