Crystallization and preliminary X-ray crystallographic analysis of the methionine sulfoxide reductase A domain of MsrAB from Haemophilus influenzae

Abstract

Methionine sulfoxide reductase (Msr) is a repair enzyme that reduces oxidized methionine to methionine. The Msr enzyme is divided into MsrA and MsrB, which reduce the S and R configurations of the substrate, respectively. In some pathogenic bacteria MsrA and MsrB exist in a fusion-protein form, MsrAB. In this study, the recombinant MsrA part of MsrAB from Haemophilus influenzae (HIMsrA) was overexpressed, purified and crystallized using the hanging-drop vapour-diffusion method. A diffraction data set was collected to 1.6 Å resolution. The crystal of HIMsrA was found to belong to space group P4(1)2(1)2, with unit-cell parameters a = b = 57.29, c = 186.28 Å, a calculated Matthews coefficient of 1.82 Å(3) Da(-1) and two molecules per asymmetric unit. A preliminary solution was determined by molecular replacement. Refinement of the structure is currently in progress.

Figure 1

SDS–PAGE analysis of purified HIMsrA (31–196). Lane M, markers (labelled in kDa); lane 1, purified HIMsrA after two-step purification.

Figure 2

Crystals of H. influenzae MsrA grown in 3.5 M sodium formate at 297 K using the hanging-drop vapour-diffusion method. The crystal dimensions are approximately 0.1 × 0.1 × 0.05 mm.

Figure 3

X-ray diffraction image of a crystal of H. influenzae MsrA. The resolution limit at the edge is 1.6 Å.