title: Blue native electrophoresis to study mitochondrial complex I in C. elegans.
abstract: 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.
authors: Department of General Pediatrics, University Children's Hospital, Heinrich-Heine-University, Düsseldorf, Germany.
authors: Daniela
authors: D
authors: van den Ecker
authors: Mariël A
authors: MA
authors: van den Brand
authors: Olaf
authors: O
authors: Bossinger
authors: Ertan
authors: E
authors: Mayatepek
authors: Leo G
authors: LG
authors: Nijtmans
authors: Felix
authors: F
authors: Distelmaier
mesh_terms: 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
chemical_list: D029742:Caenorhabditis elegans Proteins; D009243:NAD; D010088:Oxidoreductases; D042967:Electron Transport Complex I