title: Crystallization and preliminary X-ray diffraction analysis of a class II phospholipase D from Loxosceles intermedia venom.
abstract: Phospholipases D are the major dermonecrotic component of Loxosceles venom and catalyze the hydrolysis of phospholipids, resulting in the formation of lipid mediators such as ceramide-1-phosphate and lysophosphatidic acid which can induce pathological and biological responses. Phospholipases D can be classified into two classes depending on their catalytic efficiency and the presence of an additional disulfide bridge. In this work, both wild-type and H12A-mutant forms of the class II phospholipase D from L. intermedia venom were crystallized. Wild-type and H12A-mutant crystals were grown under very similar conditions using PEG 200 as a precipitant and belonged to space group P12(1)1, with unit-cell parameters a = 50.1, b = 49.5, c = 56.5 Å, β = 105.9°. Wild-type and H12A-mutant crystals diffracted to maximum resolutions of 1.95 and 1.60 Å, respectively.
authors.affiliation: Centro Multiusuário de Inovação Biomolecular, Departamento de Física, Universidade Estadual Paulista (UNESP), São José do Rio Preto-SP, Brazil.
authors.forename: Anwar
authors.initials: A
authors.lastname: Ullah
authors.forename: Priscila Oliveira
authors.initials: PO
authors.lastname: de Giuseppe
authors.forename: Mario Tyago
authors.initials: MT
authors.lastname: Murakami
authors.forename: Dilza
authors.initials: D
authors.lastname: Trevisan-Silva
authors.forename: Ana Carolina Martins
authors.initials: AC
authors.lastname: Wille
authors.forename: Daniele
authors.initials: D
authors.lastname: Chaves-Moreira
authors.forename: Luiza Helena
authors.initials: LH
authors.lastname: Gremski
authors.forename: Rafael Bertoni
authors.initials: RB
authors.lastname: da Silveira
authors.forename: Andrea
authors.initials: A
authors.lastname: Sennf-Ribeiro
authors.forename: Olga Meiri
authors.initials: OM
authors.lastname: Chaim
authors.forename: Silvio Sanches
authors.initials: SS
authors.lastname: Veiga
authors.forename: Raghuvir Krishnaswamy
authors.initials: RK
authors.lastname: Arni
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D004058:Diffusion; D004220:Disulfides / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D006639:Histidine / Q000737:chemistry; D006358:Hot Temperature; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D010739:Phospholipase D / Q000737:chemistry* / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D010727:Phosphoric Diester Hydrolases; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D013111:Spider Venoms / Q000201:enzymology*; D013112:Spiders / Q000201:enzymology*; D014169:Transformation, Bacterial; D014961:X-Ray Diffraction
chemical_list: D004220:Disulfides; D011993:Recombinant Fusion Proteins; D013111:Spider Venoms; C055161:loxosceles venom; D006639:Histidine; D010727:Phosphoric Diester Hydrolases; D010739:Phospholipase D