Crystallization and preliminary X-ray crystallographic studies of pig heart carbonyl reductase

Abstract

Pig heart carbonyl reductase (PHCR), which belongs to the short-chain dehydrogenase/reductase (SDR) family, has been crystallized by the hanging-drop vapour-diffusion method. Two crystal forms (I and II) have been obtained in the presence of NADPH. Form I crystals belong to the tetragonal space group P4(2), with unit-cell parameters a = b = 109.61, c = 94.31 A, and diffract to 1.5 A resolution. Form II crystals belong to the tetragonal space group P4(1)2(1)2, with unit-cell parameters a = b = 120.10, c = 147.00 A, and diffract to 2.2 A resolution. Both crystal forms are suitable for X-ray structure analysis at high resolution.

Figure 1

Tetragonal crystals of PHCR. (a) Form I. (b) Form II.

Figure 2

Stereographic projection of the self-rotation function in spherical polar angles at κ = 180° for the PHCR crystal. The self-rotation search was carried out with the program POLARRFN from the CCP4 suite using the data in the 20–3.0 Å resolution shell and an integration radius of 30 Å. Contour lines are drawn at an increment of 10% of the origin peak, starting at 30% of the origin peak. The program produces constant rotation angle κ for different axis directions defined by ω (angle from pole) and ϕ (angle around equator). The ϕ angles are marked on the circumference and the ω angles are defined as 0 or 180° at the centre and 90° around the edge.