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. 2008 Nov 1;64(Pt 11):1056-9.
doi: 10.1107/S1744309108032399. Epub 2008 Oct 31.

Purification, crystallization and preliminary X-ray diffraction analysis of the Whirly domain of StWhy2 in complex with single-stranded DNA

Laurent Cappadocia  1 , Jurgen Sygusch, Normand Brisson
Affiliations

Purification, crystallization and preliminary X-ray diffraction analysis of the Whirly domain of StWhy2 in complex with single-stranded DNA

Laurent Cappadocia et al. Acta Crystallogr Sect F Struct Biol Cryst Commun. .

Abstract

StWhy1 and StWhy2 are members of the Whirly family of single-stranded DNA (ssDNA) binding proteins. To understand the mode of binding of the Whirly proteins to single-stranded DNA, crystals of the Whirly domains of both StWhy1 and StWhy2 in complex with single-stranded DNA were obtained by the hanging-drop vapour-diffusion method. The diffraction patterns of the StWhy1-ssDNA complex crystals displayed severe anisotropy and were of low resolution, making them unsuitable for structure determination. In contrast, the crystals of the StWhy2-ssDNA complex diffracted isotropically to 2.20 A resolution. The crystallization and data collection to 2.20 A resolution of StWhy2 in the free form are also reported.

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Figures

Figure 1
Figure 1
(a) Crystals of StWhy1–ERE32. (b) Diffraction pattern of StWhy1–ERE32 showing high anisotropy along the c* direction (black arrow). The white arrow highlights diffraction at 3.2 Å resolution.
Figure 2
Figure 2
(a) Crystals of StWhy2–ERE32. (b) Diffraction pattern of StWhy2–ERE32 exhibiting low anisotropy.
Figure 3
Figure 3
SDS–PAGE analysis of dissolved crystals. Lane 1, purified StWhy2; lane 2, ERE32; lane 3, molecular-weight markers (kDa); lane 4, washed and dissolved crystals of StWhy2 in the free form; lane 5, washed and dissolved crystals of StWhy2–ERE32. Proteins and DNA were stained with silver nitrate.
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References

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