Structural requirements of pyrroloquinoline quinone dependent enzymatic reactions

Protein Sci. 2000 Jul;9(7):1265-73. doi: 10.1110/ps.9.7.1265.

Abstract

On the basis of crystal structures of the pyrroloquinoline quinone (PQQ) dependent enzymes methanol dehydrogenase (MDH) and soluble glucose dehydrogenase (s-GDH), different catalytic mechanisms have been proposed. However, several lines of biochemical and kinetic evidence are strikingly similar for both enzymes. To resolve this discrepancy, we have compared the structures of these enzymes in complex with their natural substrates in an attempt to bring them in line with a single reaction mechanism. In both proteins, PQQ is located in the center of the molecule near the axis of pseudo-symmetry. In spite of the absence of significant sequence homology, the overall binding of PQQ in the respective active sites is similar. Hydrogen bonding interactions are made with polar protein side chains in the plane of the cofactor, whereas hydrophobic stacking interactions are important below and above PQQ. One Arg side chain and one calcium ion are ligated to the ortho-quinone group of PQQ in an identical fashion in either active site, in agreement with their proposed catalytic function of polarizing the PQQ C5-O5 bond. The substrates are bound in a similar position above PQQ and within hydrogen bond distance of the putative general bases Asp297 (MDH) and His144 (s-GDH). On the basis of these similarities, we propose that MDH and s-GDH react with their substrates through an identical mechanism, comprising general base-catalyzed hydride transfer from the substrate to PQQ and subsequent tautomerization of the PQQ intermediate to reduced PQQ.

Publication types

  • Review

MeSH terms

  • Alcohol Oxidoreductases / chemistry
  • Alcohol Oxidoreductases / metabolism
  • Binding Sites
  • Calcium / metabolism
  • Coenzymes / chemistry
  • Coenzymes / metabolism
  • Enzymes / chemistry*
  • Enzymes / metabolism*
  • Glucose 1-Dehydrogenase
  • Glucose Dehydrogenases / chemistry
  • Glucose Dehydrogenases / metabolism
  • Models, Molecular
  • PQQ Cofactor
  • Protein Conformation
  • Quinolones / chemistry*
  • Quinolones / metabolism*
  • Quinones / chemistry*
  • Quinones / metabolism*
  • Structure-Activity Relationship

Substances

  • Coenzymes
  • Enzymes
  • Quinolones
  • Quinones
  • PQQ Cofactor
  • Alcohol Oxidoreductases
  • Glucose Dehydrogenases
  • Glucose 1-Dehydrogenase
  • alcohol dehydrogenase (acceptor)
  • Calcium