Complex III staining in blue native polyacrylamide gels.

Joél Smet; Boél De Paepe; Sara Seneca; Willy Lissens; H. Kotarsky; Linda De Meirleir; V. Fellman; Rudy Van Coster

Complex III staining in blue native polyacrylamide gels.

Joél Smet, Boél De Paepe, Sara Seneca, Willy Lissens, H. Kotarsky, Linda De Meirleir, V. Fellman, Rudy Van Coster

Research output: Contribution to journal › Article › peer-review

20 Citations (Scopus)

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 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.

Original language	English
Pages (from-to)	741-747
Number of pages <span style="color:red"p> <font size="1.5"> ✽ </span> </font>	7
Journal	J Inherit Metab Dis
Volume	34
Publication status	Published - 12 Apr 2011

Keywords

oxphos deficiencies
BN-PAGE gel
complex III staining

FWOAL377: Molecular investigations in the patients with defects in the complexes of the oxidative phosphorylation and the pyruvate dehydrogenase complex.
De Meirleir, L., Seneca, S. & Lissens, W.
1/01/06 → 31/12/09
Project: Fundamental
OZR1216: Molecular investigations in the patients with defects in the complexes of the oxidative phosphorylation and the pyruvate dehydrogenase complex.
De Meirleir, L., Lissens, W., Seneca, S. & De Meirleir, L.
1/01/06 → 31/12/08
Project: Fundamental

Cite this

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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 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.",

keywords = "oxphos deficiencies, BN-PAGE gel, complex III staining",

author = "Jo{\'e}l Smet and {De Paepe}, Bo{\'e}l and Sara Seneca and Willy Lissens and H. Kotarsky and {De Meirleir}, Linda and V. Fellman and {Van Coster}, Rudy",

year = "2011",

month = apr,

day = "12",

language = "English",

volume = "34",

pages = "741--747",

journal = "Journal of Inherited Metabolic Disease",

issn = "0141-8955",

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TY - JOUR

T1 - Complex III staining in blue native polyacrylamide gels.

AU - Smet, Joél

AU - De Paepe, Boél

AU - Seneca, Sara

AU - Lissens, Willy

AU - Kotarsky, H.

AU - De Meirleir, Linda

AU - Fellman, V.

AU - Van Coster, Rudy

PY - 2011/4/12

Y1 - 2011/4/12

N2 - 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.

AB - 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.

KW - oxphos deficiencies

KW - BN-PAGE gel

KW - complex III staining

M3 - Article

VL - 34

SP - 741

EP - 747

JO - Journal of Inherited Metabolic Disease

JF - Journal of Inherited Metabolic Disease

SN - 0141-8955

ER -

Complex III staining in blue native polyacrylamide gels.

Abstract

Keywords

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Projects

FWOAL377: Molecular investigations in the patients with defects in the complexes of the oxidative phosphorylation and the pyruvate dehydrogenase complex.

OZR1216: Molecular investigations in the patients with defects in the complexes of the oxidative phosphorylation and the pyruvate dehydrogenase complex.

Cite this