Crystallization and preliminary X-ray analysis of the reduced Rieske-type [2Fe-2S] ferredoxin derived from Pseudomonas sp. strain KKS102

Abstract

The reduced form of BphA3, a Rieske-type [2Fe-2S] ferredoxin component of the biphenyl dioxygenase BphA from Pseudomonas sp. strain KKS102, was crystallized by the sitting-drop vapour-diffusion method under anaerobic conditions. The crystal belongs to space group P3(1)21, with unit-cell parameters a = b = 49.6, c = 171.9 A, and diffracts to a resolution of 1.95 A. A molecular-replacement calculation using oxidized BphA3 as a search model yielded a satisfactory solution.

Figure 1

The anaerobic chamber used in the present study. It includes a UV–Vis spectrometer (A), a microscope with CCD (B) and an incubator for crystallization (C) in the anaerobic chamber.

Figure 2

Absorption spectrum of BphA3 in oxidized (red) and reduced (green) forms.

Figure 3

Crystals of the reduced form of BphA3 from Pseudomonas sp. strain KKS102.

Figure 4

X-ray diffraction pattern of the reduced form of the BphA3 crystal. The diffraction data were collected at beamline NW12 of PF-AR using an ADSC Quantum 210 CCD detector.