Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form

Characterization of the iron-sulfur cluster coordinated by a cysteine cluster motif (cxxcxxxcx27c) in the nqo3 subunit in the proton-translocating nadh-quinone oxidoreductase (ndh-1) of thermus thermophilus hb-8

Academic Article
uri icon
  • Overview
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

authors

  • Nakamaru-Ogiso, E.
  • Yano, T.
  • Ohnishi, T.
  • Yagi, Takao

publication date

  • 2002

journal

  • Journal of Biological Chemistry  Journal

abstract

  • The proton-translocating NADH-quinone oxidoreductase (NDH-1) of Thermus thermophilus HB-8 is composed of 14 subunits (designated Nqo1-14). This NDH-1 houses nine putative iron-sulfur binding sites, eight of which are generally found in bacterial NDH-1 and its mitochondrial counterpart (complex I). The extra site contains a CXXCXXXCX(27)C motif and is located in the Nqo3 subunit. This motif was originally found in Escherichia coli NDH-1 and was assigned to a binuclear cluster (g(z, y, x) = 2.00, 1.95, 1.92) and named N1c. In this report, the Thermus Nqo3 fragment containing this motif was heterologously overexpressed, using a glutathione S-transferase fusion system. This fragment contained a small amount of iron-sulfur cluster, whose content was significantly increased by in vitro reconstitution. The UV-visible and EPR spectroscopic properties of this fragment indicate that the ligated iron-sulfur cluster is tetranuclear with nearly axial symmetry (g( parallel, perpendicular) = 2.045, approximately 1.94). Site-directed mutants show that all four cysteines participate in the ligation of a [4Fe-4S] cluster. Considering the fact that the same motif coordinates only tetranuclear clusters in other enzymes so far known, we propose that the CXXCXXXCX(27)C motif in the Nqo3 subunit most likely ligates the [4Fe-4S] cluster.

subject areas

  • Amino Acid Sequence
  • Base Sequence
  • Cysteine
  • DNA Primers
  • Electron Spin Resonance Spectroscopy
  • Glutathione Transferase
  • Iron-Sulfur Proteins
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Protons
  • Quinone Reductases
  • Recombinant Fusion Proteins
  • Sequence Homology, Amino Acid
  • Thermus thermophilus
scroll to property group menus

Identity

International Standard Serial Number (ISSN)

  • 0021-9258

Digital Object Identifier (DOI)

  • 10.1074/jbc.M108796200

PubMed ID

  • 11704668
scroll to property group menus

Additional Document Info

start page

  • 1680

end page

  • 1688

volume

  • 277

issue

  • 3

©2019 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support