title: Disruption of individual nuo-genes leads to the formation of partially assembled NADH:ubiquinone oxidoreductase (complex I) in Escherichia coli.
abstract: The proton-pumping NADH:ubiquinone oxidoreductase, respiratory complex I, couples the electron transfer from NADH to ubiquinone with the translocation of protons across the membrane. In Escherichia coli the complex is made up of 13 different subunits encoded by the so-called nuo-genes. Mutants, in which each of the nuo-genes was individually disrupted by the insertion of a resistance cartridge were unable to assemble a functional complex I. Each disruption resulted in the loss of complex I-mediated activity and the failure to extract a structurally intact complex. Thus, all nuo-genes are required either for the assembly or the stability of a functional E. coli complex I. The three subunits comprising the soluble NADH dehydrogenase fragment of the complex were detected in the cytoplasm of several nuo-mutants as one distinct band after BN-PAGE. It is discussed that the fully assembled NADH dehydrogenase fragment represents an assembly intermediate of the E. coli complex I. A partially assembled complex I bound to the membrane was detected in the nuoK and nuoL mutants, respectively. Overproduction of the ΔNuoL variant resulted in the accumulation of two populations of a partially assembled complex in the cytoplasmic membranes. Both populations are devoid of NuoL. One population is enzymatically active, while the other is not. The inactive population is missing cluster N2 and is tightly associated with the inducible lysine decarboxylase. This article is part of a Special Issue entitled: Biogenesis/Assembly of Respiratory Enzyme Complexes.
authors: Albert-Ludwigs-Universität, Freiburg, Institut für Organische Chemie und Biochemie and Spemann Graduate School of Biology and Medicine, Albertstr. 21, 79104 Freiburg i. Br., Germany.
authors: Heiko
authors: H
authors: Erhardt
authors: Stefan
authors: S
authors: Steimle
authors: Vera
authors: V
authors: Muders
authors: Thomas
authors: T
authors: Pohl
authors: Julia
authors: J
authors: Walter
authors: Thorsten
authors: T
authors: Friedrich
mesh_terms: D002262:Carboxy-Lyases / Q000378:metabolism / Q000502:physiology; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism; D003593:Cytoplasm / Q000201:enzymology; D004578:Electron Spin Resonance Spectroscopy; D042967:Electron Transport Complex I / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D005663:Furans / Q000494:pharmacology; D017353:Gene Deletion; D015870:Gene Expression; D007506:Iron-Sulfur Proteins / Q000378:metabolism; D055503:Protein Multimerization; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D005663:Furans; D007506:Iron-Sulfur Proteins; D021122:Protein Subunits; C087673:annonin VI; D002262:Carboxy-Lyases; C021520:lysine decarboxylase; D042967:Electron Transport Complex I