Cloning, overexpression, purification, crystallization, and preliminary X-ray studies of SP_0149, the substrate binding protein of an ABC transporter from Streptococcus pneumoniae.

ABSTRACT: A truncated (29 residues from the N-terminus) and N-terminal His-tagged form of SP_0149 from pneumococcal strain ATCC BAA-334 was overexpressed and purified to homogeneity using affinity and gel-filtration chromatography. Diffraction quality crystals were grown at 293?K using the hanging-drop vapour-diffusion technique. X-ray diffraction data were collected to 2.3?Å resolution from a single-crystal that belonged to the orthorhombic space group P2(1)2(1)2(1) with the unit-cell parameters a=54.56, b=75.61, c=75.52?Å. The calculated values of the Matthews coefficient assuming one molecule (with calculated molecular weight of 30?400?Da) in the crystal asymmetric unit and the corresponding solvent content were 2.56?Å3?Da(-1) and 52.0%, respectively.

SUBMITTER: Bhushan J 

PROVIDER: S-EPMC3144799 | biostudies-literature | 2011 Jul

REPOSITORIES: biostudies-literature