title: Expression, purification and crystallization of class C acid phosphatases from Francisella tularensis and Pasteurella multocida.
abstract: Class C nonspecific acid phosphatases are bacterial enzymes that are secreted across the cytoplasmic membrane and hydrolyze a variety of phosphomonoesters at acidic pH. These enzymes are of interest for the development of improved vaccines and clinical diagnostic methods. In one case, the category A pathogen Francisella tularensis, the class C phosphatase plays a role in bacterial fitness. Here, the cloning, expression, purification and crystallization methods for the class C acid phosphatases from F. tularensis and Pasteurella multocida are reported. Crystals of the F. tularensis enzyme diffracted to 2.0 A resolution and belonged to space group C222(1), with one enzyme molecule in the asymmetric unit. Crystals of the P. multocida enzyme diffracted to 1.85 A resolution and belonged to space group C2, with three molecules in the asymmetric unit. Diffraction patterns from crystals of the P. multocida enzyme exhibited multiple interpenetrating reciprocal-space lattices, indicating epitaxial twinning. Despite this aberrance, autoindexing was robust and the data could be satisfactorily processed to 1.85 A resolution using MOSFLM and SCALA.
authors.affiliation: Department of Chemistry, University of Missouri-Columbia, 65211, USA.
authors.forename: Harkewal
authors.initials: H
authors.lastname: Singh
authors.forename: Richard L
authors.initials: RL
authors.lastname: Felts
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Malinski
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Calcutt
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Reilly
authors.forename: John J
authors.initials: JJ
authors.lastname: Tanner
mesh_terms: D000135:Acid Phosphatase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D003460:Crystallization; D018360:Crystallography, X-Ray; D005604:Francisella tularensis / Q000201:enzymology*; D016979:Pasteurella multocida / Q000201:enzymology*
chemical_list: D000135:Acid Phosphatase