References

Selected-area diffraction of a disordered polycrystalline aggregate

A. van Donkelaar, CSIRO Health Sciences


SYNOPSIS: Use of a narrow beam permits the selected-area diffraction technique to be used on a protein-crystal aggregate to isolate a good single crystal for structure refinement analysis.

The narrow focussed beam produced by the PX50 optical system permits the selected-area diffraction technique to be applied to polycrystalline aggregates in which small single protein crystals can be isolated within the 0.1mm beam diameter over a feasible oscillation range. An example of this situation, in which crystallisation of a neuraminidase mutant yielded a single polycrystalline specimen, initially yielded no useful diffraction data when irradiated centrally (see figures). The observation of a single-crystal protrusion from the aggregate prompted the attempt to orient the aggregate to isolate the single-crystal in the beam, leading to a successful structure refinement to 3Å from a crystal possessing a relatively large unit-cell volume despite an irradiated crystal volume of less than 100x40x30 mm.

Selected-area diffraction patterns from adjacent areas of a polycrystalline aggregate of neuraminidase-mutant crystals. Circles show beam position (after specimen was mounted and frozen in cryostream) corresponding to observed diffraction patterns of 40 mins exposure at 300mm camera length.
Crystal data: P2 monoclinic structure, a=315.4Å, b=67.6Å, c=92.6Å, a=90°, b=92.6°, g=90°.


Circle indicates beam size and position
graticule scale is 20 mm per small division
 

Circle indicates beam size and position
graticule scale is 20 mm per small division
 

(c) Copyright AXCO 2004

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