title: Complex III staining in blue native polyacrylamide gels.
abstract: For more than a decade now blue native polyacrylamide gel electrophoresis (BN-PAGE) has been used for the study of the oxidative phosphorylation (OXPHOS) complexes. Catalytic activities of complexes I, II, IV and V can be assessed, after separation by gel electrophoresis, by incubation of the BN-PAGE gel in specific staining solutions. However, until now, a reliable staining method for testing ubiquinol cytochrome c oxidoreductase (complex III) activity by BN-PAGE gel techniques was not available. In addition, spectrophotometric methods currently in use for detection of complex III deficiency in patients are not very sensitive. Here, we describe a newly developed diagnostic method for visualization of complex III activity by direct in-gel evaluation of ubiquinol cytochrome oxidoreductase activity. We validated the method by reporting the results in six patients with previously characterised complex III defects.
authors.affiliation: Department of Pediatrics, Division of Pediatric Neurology and Metabolism, Ghent University Hospital, De Pintelaan 185, 9000 Ghent, Belgium.
authors.forename: Joél
authors.initials: J
authors.lastname: Smet
authors.forename: Boel
authors.initials: B
authors.lastname: De Paepe
authors.forename: Sara
authors.initials: S
authors.lastname: Seneca
authors.forename: Willy
authors.initials: W
authors.lastname: Lissens
authors.forename: Heike
authors.initials: H
authors.lastname: Kotarsky
authors.forename: Linda
authors.initials: L
authors.lastname: De Meirleir
authors.forename: Vineta
authors.initials: V
authors.lastname: Fellman
authors.forename: Rudy
authors.initials: R
authors.lastname: Van Coster
mesh_terms: D000138:Acidosis / Q000378:metabolism / Q000473:pathology; D000140:Acidosis, Lactic / Q000378:metabolism / Q000473:pathology; D000180:Acrylic Resins; D016022:Case-Control Studies; D002779:Cholestasis / Q000378:metabolism / Q000473:pathology; D003116:Color; D014450:Electron Transport Complex III / Q000032:analysis / Q000737:chemistry* / Q000172:deficiency* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005317:Fetal Growth Retardation / Q000378:metabolism / Q000473:pathology; D006486:Hemosiderosis / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000378:metabolism / Q000473:pathology; D008661:Metabolism, Inborn Errors / Q000175:diagnosis / Q000378:metabolism* / Q000473:pathology; D028361:Mitochondrial Diseases / Q000151:congenital; D018482:Muscle, Skeletal / Q000737:chemistry / Q000378:metabolism / Q000473:pathology; D009206:Myocardium / Q000737:chemistry / Q000378:metabolism / Q000473:pathology; D011489:Protein Denaturation; D000608:Renal Aminoacidurias / Q000378:metabolism / Q000473:pathology; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000180:Acrylic Resins; C016680:polyacrylamide gels; D014450:Electron Transport Complex III