title: Solubilization of the NADPH-oxidase from bone marrow derived macrophages by dialkylphospholipid and purification by HPLC-anion exchange chromatography.
abstract: The microsomal NADPH-oxidase of bone marrow-derived macrophages was partially purified by HPLC-anion exchange chromatography. Particulate preparations were solubilized by the Dialkylphospholipid 2-O-methyl-1-O-octadecyl-rac-glycero-3-phosphocholine. The enzyme was enriched up to 61-fold and proved to be stable under the assay conditions. The NADPH-oxidase depended on NADPH as electron donor with a Km of 2.1 microM, whereas no O2- -production was detected with up to 40 microM NADH. Ca2+ was found to be a strong inhibitor of the enzyme, whereas Mg2+ had no influence. O2- was determined by the very sensitive and selective method of superoxide dismutase-inhibitable chemiluminescence of lucigenin.
authors.affiliation: Max-Planck-Institut für Immunobiologie, Freiburg.
authors.forename: J
authors.initials: J
authors.lastname: Storch
authors.forename: E
authors.initials: E
authors.lastname: Ferber
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005260:Female; D007700:Kinetics; D008264:Macrophages / Q000201:enzymology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009247:NADH, NADPH Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism; D019255:NADPH Oxidases; D010743:Phospholipids; D012995:Solubility
chemical_list: D010743:Phospholipids; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases