Blue native electrophoresis to study mitochondrial complex I in C. elegans.
Blue native polyacrylamide gel electrophoresis (BN-PAGE) is an essential tool for investigating mitochondrial respiratory chain complexes. However, with current BN-PAGE protocols for Caenorhabditis elegans (C. elegans), large worm amounts and high quantities of mitochondrial protein are required to yield clear results. Here, we present an efficient approach to isolate mitochondrial complex I (NADH:ubiquinone oxidoreductase) from C. elegans, grown on agar plates. We demonstrate that considerably lower amounts of mitochondrial protein are sufficient to isolate complex I and to display clear in-gel activity results. Moreover, we present the first complex I assembly profile for C. elegans, obtained by two-dimensional BN/SDS-PAGE.
van den Ecker
Daniela
D
Department of General Pediatrics, University Children's Hospital, Heinrich-Heine-University, Düsseldorf, Germany.
van den Brand
Mariël A
MA
Bossinger
Olaf
O
Mayatepek
Ertan
E
Nijtmans
Leo G
LG
Distelmaier
Felix
F
D000818:Animals; D017173:Caenorhabditis elegans / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D042967:Electron Transport Complex I / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008928:Mitochondria / Q000378:metabolism*; D009243:NAD / Q000737:chemistry; D010088:Oxidoreductases / Q000737:chemistry
D029742:Caenorhabditis elegans Proteins; D009243:NAD; D010088:Oxidoreductases; D042967:Electron Transport Complex I