title: Crystallization, optimization and preliminary X-ray characterization of a metal-dependent PI-PLC from Streptomyces antibioticus.
abstract: A recombinant metal-dependent phosphatidylinositol-specific phospholipase C (PI-PLC) from Streptomyces antibioticus has been crystallized by the hanging-drop method with and without heavy metals. The native crystals belonged to the orthorhombic space group P222, with unit-cell parameters a=41.26, b=51.86, c=154.78 Å. The X-ray diffraction results showed significant differences in the crystal quality of samples soaked with heavy atoms. Additionally, drop pinning, which increases the surface area of the drops, was also used to improve crystal growth and quality. The combination of heavy-metal soaks and drop pinning was found to be critical for producing high-quality crystals that diffracted to 1.23 Å resolution.
authors.affiliation: Department of Chemistry, Bucknell University, Lewisburg, PA 17837, USA.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Jackson
authors.forename: Thomas L
authors.initials: TL
authors.lastname: Selby
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002104:Cadmium / Q000737:chemistry; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D007495:Iridium / Q000737:chemistry; D054801:Phosphoinositide Phospholipase C / Q000737:chemistry* / Q000302:isolation & purification; D013303:Streptomyces antibioticus / Q000201:enzymology*; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D002104:Cadmium; D007495:Iridium; D054801:Phosphoinositide Phospholipase C