Transmembrane orientation and topology of the NADH:quinone oxidoreductase putative quinone binding subunit NuoH

Research output: Contribution to journalArticle

Abstract

NADH:quinone oxidoreductase, or Complex I, is a multi-subunit membrane-bound enzyme in the respiratory chain of many pro- and eukaryotes. The enzyme catalyzes the oxidation of NADH and donates electrons to the quinone pool, coupled to proton translocation across the membrane, but the mechanism of energy transduction is not understood. In bacteria the enzyme consists of 14 subunits, seven membrane spanning and seven protruding from the membrane. The hydrophobic NuoH (NQO8, ND1, NAD1, NdhA) subunit is seemingly involved in quinone binding. A homologous, structurally and most likely functionally similar subunit is also found in F420H2 oxidoreductases and in complex membrane-bound hydrogenases. We have made theoretical analyses of NuoH and NuoH-like polypeptides and experimentally analyzed the transmembrane topology of the NuoH subunit from Rhodobacter capsulatus by constructing and analyzing alkaline phosphatase fusion proteins. This demonstrated that the NuoH polypeptide has eight transmembrane segments, and four highly conserved hydrophilic sequence motifs facing the inside, bacterial cytoplasm. The N-terminal and C-terminal ends are located on the outside of the membrane. A topology model of NuoH based on these results is presented, and implications from the model are discussed.

Details

Authors
  • Robert Roth
  • Cecilia Hägerhäll
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biological Sciences

Keywords

  • Complex I, NADH:quinone oxidoreductase, Hydrogenase, F420, NuoH, ND1, NQO8, NAD1, NdhA, Alkaline phosphatase fusion, (Rhodobacter capsulatus)
Original languageEnglish
Pages (from-to)352-362
JournalBiochimica et Biophysica Acta - Bioenergetics
Volume1504
Issue number2-3
Publication statusPublished - 2001
Publication categoryResearch
Peer-reviewedYes