tumor in the light of the revised SIOP-01 classification

Annals of Diagnostic Paediatric Pathology 2006, 10 (3–4): 89–92
© Copyright by Polish Paediatric Pathology Society Annals of
Blue Native Electrophoresis: an additional useful tool to study
deficiencies of mitochondrial respiratory chain complexes
Agnieszka Karkuciñska-Wiêckowska 1 , Katarzyna Czajka 2 , Micha³ Wasilewski 2 ,
Jolanta Sykut-Cegielska 3 , Maciej Pronicki 1 , Bo¿ena Cukrowska 1 , Ewa Pronicka 3 ,
Krzysztof Zab³ocki 2 , Jerzy Duszyñski 2 , Mariusz R. Wiêckowski 2
1 Department of Pathology
3 Division of Metabolic Diseases, Department of Pediatric
The Children's Memorial Health Institute
Warsaw, Poland
2 Department of Cellular Biochemistry
Nencki Institute of Experimental Biology, Polish Academy of Sciences
Warsaw, Poland
Introduction
Abstract
Blue Native polyacrylamide gel electrophoresis (BN-PAGE),
originally described by Schägger and von Jagow in 1991 [4],
is an elegant method to study protein complexes from mitochondrial
membranes. It is important that BN-PAGE enables
separation of membrane protein complexes from organelles
Address for correspondence
Several mitochondrial disorders are connected with the decreased activity of the respiratory chain which
leads to disturbance in cell metabolism and eventually to severe pathologic changes in the organism.
Localization and measurement of changes in the amount and activity of respiratory chain complexes can
provide important information of the disease etiology. Components of the mitochondrial respiratory chain,
located in the inner mitochondrial membrane and grouped in four complexes (additionally, mitochondrial
ATPase is called – complex V), are working together providing a pathway for redox processes. A wide
spectrum of methods enables studying the activity of respiratory chain complexes in tissues, cell cultures
and in isolated mitochondria. Under special experimental conditions the activity of each complex can be
measured separately using spectrophotometric methods. Acrylamide Blue Native Electrophoresis,
commonly used in molecular biology to study composition of protein complexes and protein-protein
interactions, can be successfully adapted for diagnosis of mitochondrial diseases connected with
abnormalities of the respiratory chain. BN-PAGE and in-gel activity assay can be successfully applied either
for detection of respiratory chain complex deficiency or for visualization deficiencies of individual complex
activities.
Key words: Blue Native Electrophoresis, mitochondria, respiratory chain complexes, mitochondrial disorders
Diagnostic
Paediatric
Pathology
or membrane fractions, keeping their properties and enzymatic
activities unchanged. BN-PAGE, supplemented by
other methods, e.g., in gel activity assay, spectrophotometric
measurement of the mitochondrial respiratory chain activity
and SDS-PAGE (as a second dimension), can be successfully
used in diagnosis of respiratory chain complex deficiencies.
Mariusz R. Wiêckowski 3 Pasteur Street, 02-093 Warsaw, Poland
Laboratory of Bioenergetics, Biomembranes and Metabolic Regulation tel. +48 22 589-23-72
Department of Cellular Biochemistry fax +48 22 822-53-42
Nencki Institute of Experimental Biology e-mail: m.wieckowski@nencki.gov.pl
Polish Academy of Sciences