Browsing by Subject "NQR"

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Powerful proteins
structure and function of catalytic subunits of electrogenic NADH:quinone oxidoreductases
(2013) Steffen, Wojtek; Fritz-Steuber, Julia
Electrogenic NADH:quinone oxidoreductases are large, membrane-embedded enzyme complexes found in the respiratory chain of prokaryotes and the mitochondria of eukaryotes. They represent the first module of the oxidative phosphorylation system which converts the energy from nutrients into an electrochemical gradient by coupling redox reactions to the translocation of cations across membranes. A long chain of events, such as the synthesis of ATP, ion homeostasis, reactive oxygen species production and bacterial motility depend on the activity of these complexes. Complex I consists of up to 45 subunits and can be found in the inner mitochondrial membrane of eukaryotes and in prokaryotes, where it is called NDH I. We investigated the isolated, hydrophobic ND5 subunit, which shows homologies to cation/proton antiporters, from human or Yarrowia lipolytica complex I. In vivo and biochemical analyses provided data on the cation translocation function and the alteration of function by disease-associated mutations. Taken together with the recently published 3D structure of bacterial complex I, these data allowed us to demonstrate that the ND5 subunit could possibly act as an antiporter module of mitochondrial complex I. Sodium ion translocating NADH:quinone oxidoreductase (Na+-NQR) is an enzyme found in many pathogenic bacteria. It consists of six subunits (NqrA - NqrF) whose 3D structures and enzymatic mechanisms were not known in detail at the time this project was initiated. By using high-resolution X-ray structures and site-directed mutagenesis, combined with biochemical studies, we proposed a model for catalysis and substrate selectivity on the atomic level of the electron input module of the complex, the NADH oxidizing domain of subunit NqrF. Furthermore, we analyzed the binding of silver ions to a cysteine residue in the NADH binding pocket and found that it leads to the inhibition of the Na+-NQR and to the killing of Vibrio cholerae in the nanomolar range. Subunit NqrA forms part of the quinone reductase module. By the use of physicochemical and biochemical methods we identified the herbicide 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB) as a quinone antagonist and inhibitor of the Na+-NQR complex and discovered two adjacent quinone binding sites on NqrA.
The RNF/NQR redox pumps: a versatile system for energy transduction in bacteria and archaea
(2025) Buckel, Wolfgang; Ermler, Ulrich; Vonck, Janet; Fritz, Günter; Steuber, Julia
The Na + (or H + )-translocating ferredoxin:NAD + oxidoreductase (also called RNF, rhodobacter nitrogen fixation, complex) catalyzes the oxidation of reduced ferredoxin with NAD + , hereby generating an electrochemical gradient. In the reverse reaction driven by an electrochemical gradient, RNF provides reduced ferredoxin using NADH as electron donor. RNF plays a crucial role in the metabolism of many anaerobes, such as amino acid fermenters, acetogens, or aceticlastic methanogens. The Na + -translocating NADH:quinone oxidoreductase (NQR), which has evolved from an RNF, is found in selected bacterial groups including anaerobic, marine, or pathogenic organisms. Since NQR and RNF are not related to eukaryotic respiratory complex I (NADH:quinone oxidoreductase), members of this oxidoreductase family are promising targets for novel antibiotics. RNF and NQR share a membrane-bound core complex consisting of four subunits, which represent an essential functional module for redox-driven cation transport. Several recent 3D structures of RNF and NQR in different states put forward conformational coupling of electron transfer and Na + translocation reaction steps. Based on this common principle, putative reaction mechanisms of RNF and NQR redox pumps are compared. Key points: • Electrogenic ferredoxin:NAD + oxidoreductases (RNF complexes) are found in bacteria and archaea. • The Na + -translocating NADH:quinone oxidoreductase (NQR) is evolutionary related to RNF. • The mechanism of energy conversion by RNF/NQR complexes is based on conformational coupling of electron transfer and cation transport reactions.