[Application of "blue native" electrophoresis in the studies of mitochondrial respiratory chain complexes in physiology and pathology].
"Blue Native" polyacrylamide gel electrophoresis (BN-PAGE), originally described by Schägger and von Jagow in 1991, is an elegant method to study protein complexes from mitochondrial membranes. BN-PAGE, commonly used in molecular biology to study composition of protein complexes and protein-protein interactions, enables separation of respiratory chain complexes keeping their properties and enzymatic activities unchanged. BN-PAGE, supplemented by other methods, e.g. in gel activity assay, SDS-PAGE (as a first or second dimension) can be successfully adapted for diagnosis of mitochondrial diseases connected with abnormalities of the respiratory chain. Therefore, to make a correct diagnosis of the deficiency of respiratory chain complexes, other methods, as histochemical colorimetric reactions allowing evaluation of the OXPHOS catalytic activity in individual cells and spectrophotometric technique should be used simultaneously with BN-PAGE.
Lebiedzińska
Magdalena
M
Pracownia Bioenergetyki i Blon Biologicznych, Instytut Biologii Doświadczalnej PAN im. Marcelego Nenckiego, Warszawa.
Duszyński
Jerzy
J
Wieckowski
Mariusz R
MR
D003576:Electron Transport Complex IV / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D008928:Mitochondria / Q000737:chemistry* / Q000378:metabolism*; D028361:Mitochondrial Diseases / Q000175:diagnosis / Q000473:pathology; D024101:Mitochondrial Proteins / Q000032:analysis; D046912:Multiprotein Complexes / Q000032:analysis; D012394:Rosaniline Dyes / Q000032:analysis* / Q000737:chemistry; D012995:Solubility; D013194:Staining and Labeling / Q000379:methods*
D024101:Mitochondrial Proteins; D046912:Multiprotein Complexes; D012394:Rosaniline Dyes; D003576:Electron Transport Complex IV; C004692:coomassie Brilliant Blue