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.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Crystallization
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Crystallography, X-Ray / methods
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Diffusion
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Disulfides / chemistry
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Escherichia coli / genetics
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Histidine / chemistry
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Hot Temperature
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Hydrogen Bonding
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Hydrogen-Ion Concentration
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Molecular Sequence Data
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Molecular Weight
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Mutation
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Phospholipase D / chemistry*
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Phospholipase D / classification*
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Phospholipase D / genetics
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Phospholipase D / isolation & purification
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Phosphoric Diester Hydrolases
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / classification
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Recombinant Fusion Proteins / isolation & purification
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Sequence Homology, Amino Acid
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Spider Venoms / enzymology*
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Spiders / enzymology*
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Transformation, Bacterial
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X-Ray Diffraction
Substances
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Disulfides
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Recombinant Fusion Proteins
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Spider Venoms
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loxosceles venom
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Histidine
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Phosphoric Diester Hydrolases
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Phospholipase D