title: Expression, purification and crystallization of an atypical class C acid phosphatase from Mycoplasma bovis.
abstract: Class C acid phosphatases (CCAPs) are 25-30 kDa bacterial surface proteins that are thought to function as broad-specificity 5',3'-nucleotidases. Analysis of the newly published complete genome sequence of Mycoplasma bovis PG45 revealed a putative CCAP with a molecular weight of 49.9 kDa. The expression, purification and crystallization of this new family member are described here. Standard purification procedures involving immobilized metal-ion affinity chromatography and ion-exchange chromatography yielded highly pure and crystallizable protein. Crystals were grown in sitting drops at room temperature in the presence of PEG 3350 and HEPES buffer pH 7.5 and diffracted to 2.3 Å resolution. Analysis of diffraction data suggested a primitive monoclinic space group, with unit-cell parameters a = 78, b = 101, c = 180 Å, β = 92°. The asymmetric unit is predicted to contain six molecules, which are likely to be arranged as three dimers.
authors.affiliation: Department of Chemistry, University of Missouri-Columbia, Columbia, MO 65211, USA.
authors.forename: Harkewal
authors.initials: H
authors.lastname: Singh
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Reilly
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Calcutt
authors.forename: John J
authors.initials: JJ
authors.lastname: Tanner
mesh_terms: D000135:Acid Phosphatase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D000595:Amino Acid Sequence; D003460:Crystallization; D015870:Gene Expression; D008969:Molecular Sequence Data; D045682:Mycoplasma bovis / Q000201:enzymology*; D016415:Sequence Alignment
chemical_list: D000135:Acid Phosphatase
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