Use of electron microscopy in the examination of lattice defects in crystals of alcohol oxidase

FEBS Lett. 1989 Feb 13;244(1):213-6. doi: 10.1016/0014-5793(89)81195-0.

Abstract

Alcohol oxidase, purified from the yeast Hansenula polymorpha, was crystallized in vitro for the purpose of determining its structure at atomic resolution by X-ray diffraction methods. The crystals obtained yielded only extremely weak diffraction patterns: the maximal resolution observed was in the best case 6 A. Electron microscopy of thin sections indicated that most crystals showed lattice defects which might explain the poor diffraction patterns: most surprising was the appearance of large holes interrupting an otherwise regular lattice in one of the crystal forms examined. Our results indicate that transmission electron microscopy is a suitable tool for the inspection of crystals to be used in X-ray crystallography. The method allows rapid determination of lattice defects and enables optimization of crystallization conditions.

MeSH terms

  • Alcohol Oxidoreductases*
  • Calcium Chloride
  • Crystallization
  • Dithiothreitol
  • Edetic Acid
  • Macromolecular Substances
  • Microscopy, Electron
  • Molecular Weight
  • Pichia / enzymology*
  • Saccharomycetales / enzymology*
  • X-Ray Diffraction

Substances

  • Macromolecular Substances
  • Edetic Acid
  • Alcohol Oxidoreductases
  • alcohol oxidase
  • Calcium Chloride
  • Dithiothreitol