Crystallization and preliminary X-ray analysis of PH1010 from Pyrococcus horikoshii OT3, a member of the archaeal DUF54 family of proteins

Abstract

PH1010 from Pyrococcus horikoshii OT3, a member of the archaeal DUF54 family of proteins, was expressed, purified and crystallized. Crystallization was performed by the sitting-drop vapour-diffusion method using PEG 3350 as the precipitant. The crystal diffracted X-rays to 1.90 A resolution using a synchrotron-radiation source. The space group of the crystal was determined to be P2(1)2(1)2(1), with unit-cell parameters a = 46.9, b = 49.5, c = 132.7 A. The crystal contained two PH1010 molecules in the asymmetric unit (V(M) = 2.4 A(3) Da(-1)) and had a solvent content of 48%.

Figure 1

SDS–PAGE of the purified PH1010. Lane M, molecular-weight markers (kDa). Lane 1, PH1010.

Figure 2

Images of typical PH1010 crystals. (a) Crystals of the native protein of approximate dimensions 0.25 × 0.1 × 0.05 mm. (b) Crystals of the SeMet derivative of approximate dimensions 0.25 × 0.1 × 0.05 mm.

Figure 3

An X-ray diffraction image from a typical crystal of native protein. Diffraction data are detectable to 1.90 Å. The edge of the detector corresponds to a resolution of 1.90 Å.