, Volume 34, Issue 3, pp 741-747
Date: 12 Apr 2011

Complex III staining in blue native polyacrylamide gels

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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 electroforesis, 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.

Communicated by: John Christodoulou.
References to electronic databases: Mitochondrial complex III deficiency: OMIM 124000. UQCRQ encoding subunit 7: OMIM 61280. UQCRB encoding ubiquinol-cytochrome c reductase binding protein: OMIM 191330. Cytochrome b of Complex IIII, MTCYB: OMIM 516020. GRACILE syndrome (Growth retardation, Amino aciduria, Cholestasis, Iron overload, Lactic acidosis): OMIM 603358. Bjornstad syndrome: OMIM 262000. NADH:ubiquinone oxidoreductase (complex I): EC Succinate:ubiquinone oxidoreductase (complex II): EC Ubiquinol:ferricytochrome-c oxidoreductase (complex III): EC Ferrocytochrome-c:oxygen oxidoreductase (complex IV): EC ATP phosphohydrolase (complex V): EC Deoxynucleoside-triphosphate:DNA deoxynucleotidyltransferase (DNA-directed; DNA polymerase gamma): EC Mitochondrial thymidine kinase 2: EC
Competing of interest: None declared.