Characterisation of the PQQ cofactor radical in quinoprotein ethanol dehydrogenase of Pseudomonas aeruginosa by electron paramagnetic resonance spectroscopy

FEBS Lett. 2004 Apr 23;564(1-2):69-72. doi: 10.1016/S0014-5793(04)00317-5.

Abstract

The binding pocket of the pyrroloquinoline quinone (PQQ) cofactor in quinoprotein alcohol dehydrogenases contains a characteristic disulphide ring formed by two adjacent cysteine residues. To analyse the function of this unusual structural motif we have investigated the wild-type and a double cysteine:alanine mutant of the quinoprotein ethanol dehydrogenase from Pseudomonas aeruginosa by electron paramagnetic resonance (EPR) spectroscopy. Thus, we have obtained the principal values for the full rhombic g-tensor of the PQQ semiquinone radical by high-field (94 GHz) EPR necessary for a discrimination of radical species in dehydrogenases containing PQQ together with other redox-active cofactors. Our results show that the characteristic disulphide ring is no prerequisite for the formation of the functionally important semiquinone form of PQQ.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Oxidoreductases / chemistry*
  • Alcohol Oxidoreductases / genetics
  • Bacterial Proteins / chemistry
  • Binding Sites
  • Cysteine
  • Disulfides
  • Electron Spin Resonance Spectroscopy*
  • Mutagenesis, Site-Directed
  • Oxidation-Reduction
  • PQQ Cofactor / chemistry*
  • Pseudomonas aeruginosa / enzymology*

Substances

  • Bacterial Proteins
  • Disulfides
  • PQQ Cofactor
  • Alcohol Oxidoreductases
  • alcohol dehydrogenase (acceptor)
  • Cysteine