Preliminary X-ray crystallographic analysis of SMU.2055 protein from the caries pathogen Streptococcus mutans.

Abstract

The SMU.2055 gene from the major caries pathogen Streptococcus mutans is annotated as a putative acetyltransferase with 163 amino-acid residues. In order to identify its function via structural studies, the SMU.2055 gene was cloned into the expression vector pET28a. Native and SeMet-labelled SMU.2055 proteins with a His(6) tag at the N-terminus were expressed at a high level in Escherichia coli strain BL21 (DE3) and purified to homogeneity by Ni(2+)-chelating affinity chromatography. Diffraction-quality crystals of SeMet-labelled SMU.2055 were obtained using the sitting-drop vapour-diffusion method and diffracted to a resolution of 2.5 A on beamline BL17A at the Photon Factory, Tsukuba, Japan. The crystals belong to the orthorhombic space group C222(1), with unit-cell parameters a = 92.0, b = 95.0, c = 192.2 A. The asymmetric unit contained four molecules, with a solvent content of 57.1%.