Comparison of the decameric structure of peroxiredoxin-II by transmission electron microscopy and X-ray crystallography

Biochim Biophys Acta. 2001 Jun 11;1547(2):221-34. doi: 10.1016/s0167-4838(01)00184-4.

Abstract

The decameric human erythrocyte protein torin is identical to the thiol-specific antioxidant protein-II (TSA-II), also termed peroxiredoxin-II (Prx-II). Single particle analysis from electron micrographs of Prx-II molecules homogeneously orientated across holes in the presence of a thin film of ammonium molybdate and trehalose has facilitated the production of a >/=20 A 3-D reconstruction by angular reconstitution that emphasises the D5 symmetry of the ring-like decamer. The X-ray structure for Prx-II was fitted into the transmission electron microscopic reconstruction by molecular replacement. The surface-rendered transmission electron microscopy (TEM) reconstruction correlates well with the solvent-excluded surface of the X-ray structure of the Prx-II molecule. This provides confirmation that transmission electron microscopy of negatively stained specimens, despite limited resolution, has the potential to reveal a valid representation of surface features of protein molecules. 2-D crystallisation of the Prx-II protein on mica as part of a TEM study resulted in the formation of a p2 crystal form with parallel linear arrays of stacked rings. This latter 2-D form correlates well with that observed from the 2.7 A X-ray structure of Prx-II solved from a new orthorhombic 3-D crystal form.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallography, X-Ray
  • Erythrocytes / chemistry
  • Humans
  • Microscopy, Electron
  • Models, Molecular
  • Molybdenum
  • Peroxidases / chemistry*
  • Peroxidases / isolation & purification
  • Peroxidases / ultrastructure
  • Peroxiredoxins
  • Surface Properties
  • Trehalose

Substances

  • Molybdenum
  • Trehalose
  • Peroxidases
  • Peroxiredoxins
  • ammonium molybdate