Tanaka_2018_Acta.Crystallogr.F.Struct.Biol.Commun_74_567

Staphylococcus aureus lipase (SAL), a triacylglycerol esterase, is an important virulence factor in S. aureus and may be a therapeutic target for infectious diseases caused by S. aureus. For the purposes of anti-SAL drug development using structure-based drug design, X-ray crystallographic analysis of SAL overexpressed in Escherichia coli was performed. The recombinant protein was purified using a three-step protocol involving immobilized metal-affinity chromatography, cation-exchange chromatography and anion-exchange chromatography flowthrough methods, yielding 40 mg of protein per litre of bacterial culture. Crystals were obtained using the sitting-drop vapor-diffusion technique. Diffraction data to 3.0 A resolution were collected on the BL44XU beamline at SPring-8 at the zinc peak of 1.2842 A for SAD phasing. The crystals belonged to space group P4122 or P4322, with unit-cell parameters a = 131.0, b = 131.0, c = 250.6 A, and are likely to contain four SAL molecules (408 residues) per asymmetric unit.