title: Exploring the catalytic core of complex I by Yarrowia lipolytica yeast genetics.
abstract: We have developed Yarrowia lipolytica as a model system to study mitochondrial complex I that combines the application of fast and convenient yeast genetics with efficient structural and functional analysis of its very stable complex I isolated by his-tag affinity purification with high yield. Guided by a structural model based on homologies between complex I and [NiFe] hydrogenases mutational analysis revealed that the 49 kDa subunit plays a central functional role in complex I. We propose that critical parts of the catalytic core of complex I have evolved from the hydrogen reactive site of [NiFe] hydrogenases and that iron-sulfur cluster N2 resides at the interface between the 49 kDa and PSST subunits. These findings are in full agreement with the "semiquinone switch" mechanism according to which coupling of electron and proton transfer in complex I is achieved by a single integrated pump comprising cluster N2, the binding site for substrate ubiquinone, and a tightly bound quinone or quinoid group.
authors.affiliation: Universitätsklinikum Frankfurt, Institut für Biochemie I, Zentrum der Biologischen Chemie, Frankfurt am Main, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Kerscher
authors.forename: N
authors.initials: N
authors.lastname: Kashani-Poor
authors.forename: K
authors.initials: K
authors.lastname: Zwicker
authors.forename: V
authors.initials: V
authors.lastname: Zickermann
authors.forename: U
authors.initials: U
authors.lastname: Brandt
mesh_terms: D000595:Amino Acid Sequence; D020134:Catalytic Domain / Q000235:genetics; D042967:Electron Transport Complex I; D005800:Genes, Fungal; D008928:Mitochondria / Q000201:enzymology; D008957:Models, Genetic; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D021122:Protein Subunits; D017386:Sequence Homology, Amino Acid; D025062:Yarrowia / Q000201:enzymology* / Q000235:genetics*
chemical_list: D021122:Protein Subunits; D009247:NADH, NADPH Oxidoreductases; D042967:Electron Transport Complex I