Identification of the ubiquinone-binding site of NADH:ubiquinone oxidoreductase (complex I) from Neurospora crassa

Biochemistry ◽  
1992 ◽  
Vol 31 (46) ◽  
pp. 11413-11419 ◽  
Author(s):  
Helga Heinrich ◽  
Sigurd Werner
Keyword(s):  
Neurospora Crassa ◽  
Binding Site ◽  
Complex I ◽  
Nadh:Ubiquinone Oxidoreductase ◽  
Ubiquinone Binding

scholarly journals Accessory NUMM (NDUFS6) subunit harbors a Zn-binding site and is essential for biogenesis of mitochondrial complex I

2015 ◽  
Vol 112 (18) ◽  
pp. 5685-5690 ◽  
Author(s):  
Katarzyna Kmita ◽  
Christophe Wirth ◽  
Judith Warnau ◽  
Sergio Guerrero-Castillo ◽  
Carola Hunte ◽  
...  
Keyword(s):  
Binding Site ◽  
Complex I ◽  
Proton Pumping ◽  
Nadh:Ubiquinone Oxidoreductase ◽  
X Ray Crystallography ◽  
Late Step ◽  
Multistep Process ◽  
Assembly Factor ◽  
Accessory Subunit ◽  
Assembly Intermediate

Mitochondrial proton-pumping NADH:ubiquinone oxidoreductase (respiratory complex I) comprises more than 40 polypeptides and contains eight canonical FeS clusters. The integration of subunits and insertion of cofactors into the nascent complex is a complicated multistep process that is aided by assembly factors. We show that the accessory NUMM subunit of complex I (human NDUFS6) harbors a Zn-binding site and resolve its position by X-ray crystallography. Chromosomal deletion of the NUMM gene or mutation of Zn-binding residues blocked a late step of complex I assembly. An accumulating assembly intermediate lacked accessory subunit N7BM (NDUFA12), whereas a paralog of this subunit, the assembly factor N7BML (NDUFAF2), was found firmly bound instead. EPR spectroscopic analysis and metal content determination after chromatographic purification of the assembly intermediate showed that NUMM is required for insertion or stabilization of FeS cluster N4.

scholarly journals Inhibitors of ROS production by the ubiquinone-binding site of mitochondrial complex I identified by chemical screening

2013 ◽  
Vol 65 ◽  
pp. 1047-1059 ◽  
Author(s):  
Adam L. Orr ◽  
Deepthi Ashok ◽  
Melissa R. Sarantos ◽  
Tong Shi ◽  
Robert E. Hughes ◽  
...  
Keyword(s):  
Binding Site ◽  
Complex I ◽  
Mitochondrial Complex I ◽  
Ros Production ◽  
Mitochondrial Complex ◽  
Chemical Screening ◽  
Ubiquinone Binding

scholarly journals Structure of inhibitor-bound mammalian complex I

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Hannah R. Bridges ◽  
Justin G. Fedor ◽  
James N. Blaza ◽  
Andrea Di Luca ◽  
Alexander Jussupow ◽  
...  
Keyword(s):  
Complex I ◽  
Energy Coupling ◽  
Competitive Inhibitor ◽  
Nadh:Ubiquinone Oxidoreductase ◽  
Mouse Heart ◽  
Coupling Mechanism ◽  
Substrate Reduction ◽  
Mitochondrial Inner Membrane ◽  
Respiratory Complex I ◽  
Ubiquinone Binding

Abstract Respiratory complex I (NADH:ubiquinone oxidoreductase) captures the free energy from oxidising NADH and reducing ubiquinone to drive protons across the mitochondrial inner membrane and power oxidative phosphorylation. Recent cryo-EM analyses have produced near-complete models of the mammalian complex, but leave the molecular principles of its long-range energy coupling mechanism open to debate. Here, we describe the 3.0-Å resolution cryo-EM structure of complex I from mouse heart mitochondria with a substrate-like inhibitor, piericidin A, bound in the ubiquinone-binding active site. We combine our structural analyses with both functional and computational studies to demonstrate competitive inhibitor binding poses and provide evidence that two inhibitor molecules bind end-to-end in the long substrate binding channel. Our findings reveal information about the mechanisms of inhibition and substrate reduction that are central for understanding the principles of energy transduction in mammalian complex I.

scholarly journals Supramolecular Organization of the Respiratory Chain in Neurospora crassa Mitochondria

2007 ◽  
Vol 6 (12) ◽  
pp. 2391-2405 ◽  
Author(s):  
Isabel Marques ◽  
Norbert A. Dencher ◽  
Arnaldo Videira ◽  
Frank Krause
Keyword(s):  
Neurospora Crassa ◽  
Respiratory Chain ◽  
Complex I ◽  
Polyacrylamide Gel Electrophoresis ◽  
Nadh:Ubiquinone Oxidoreductase ◽  
Deletion Mutants ◽  
Supramolecular Organization ◽  
Wild Type ◽  
Respiratory Supercomplexes ◽  
Native Polyacrylamide Gel Electrophoresis

ABSTRACT The existence of specific respiratory supercomplexes in mitochondria of most organisms has gained much momentum. However, its functional significance is still poorly understood. The availability of many deletion mutants in complex I (NADH:ubiquinone oxidoreductase) of Neurospora crassa, distinctly affected in the assembly process, offers unique opportunities to analyze the biogenesis of respiratory supercomplexes. Herein, we describe the role of complex I in assembly of respiratory complexes and supercomplexes as suggested by blue and colorless native polyacrylamide gel electrophoresis and mass spectrometry analyses of mildly solubilized mitochondria from the wild type and eight deletion mutants. As an important refinement of the fungal respirasome model, we found that the standard respiratory chain of N. crassa comprises putative complex I dimers in addition to I-III-IV and III-IV supercomplexes. Three Neurospora mutants able to assemble a complete complex I, lacking only the disrupted subunit, have respiratory supercomplexes, in particular I-III-IV supercomplexes and complex I dimers, like the wild-type strain. Furthermore, we were able to detect the I-III-IV supercomplexes in the nuo51 mutant with no overall enzymatic activity, representing the first example of inactive respirasomes. In addition, III-IV supercomplexes were also present in strains lacking an assembled complex I, namely, in four membrane arm subunit mutants as well as in the peripheral arm nuo30.4 mutant. In membrane arm mutants, high-molecular-mass species of the 30.4-kDa peripheral arm subunit comigrating with III-IV supercomplexes and/or the prohibitin complex were detected. The data presented herein suggest that the biogenesis of complex I is linked with its assembly into supercomplexes.

scholarly journals The Ubiquinone-binding Site in NADH:Ubiquinone Oxidoreductase fromEscherichia coli

2003 ◽  
Vol 278 (28) ◽  
pp. 25731-25737 ◽  
Author(s):  
Xing Gong ◽  
Tong Xie ◽  
Linda Yu ◽  
Micaela Hesterberg ◽  
Dierk Scheide ◽  
...  
Keyword(s):  
Binding Site ◽  
Nadh:Ubiquinone Oxidoreductase ◽  
Fromescherichia Coli ◽  
Ubiquinone Binding
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