Use of electron microscopy in the examination of lattice defects in crystals of alcohol oxidase.
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.
Van der Klei
I J
IJ
Department of Microbiology, University of Groningen, The Netherlands.
Lawson
C L
CL
Rozeboom
H
H
Dijkstra
B W
BW
Veenhuis
M
M
Harder
W
W
Hol
W G
WG
D000429:Alcohol Oxidoreductases; D002122:Calcium Chloride; D003460:Crystallization; D004229:Dithiothreitol; D004492:Edetic Acid; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008970:Molecular Weight; D010843:Pichia / Q000201:enzymology*; D004718:Saccharomycetales / Q000201:enzymology*; D014961:X-Ray Diffraction
D046911:Macromolecular Substances; D004492:Edetic Acid; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D002122:Calcium Chloride; D004229:Dithiothreitol