Crystallization and preliminary X-ray diffraction analysis of human phosphate-binding protein

Xanthomonas axonopodispv.citri(X. citri) is an important bacterium that causes citrus canker disease in plants in Brazil and around the world, leading to significant economic losses. Determination of the physiology and mechanisms of pathogenesis of this bacterium is an important step in the development of strategies for its containment. Phosphate is an essential ion in all microrganisms owing its importance during the synthesis of macromolecules and in gene and protein regulation. Interestingly,X. citrihas been identified to present two periplasmic binding proteins that have not been further characterized: PstS, from an ATP-binding cassette for high-affinity uptake and transport of phosphate, and PhoX, which is encoded by an operon that also contains a putative porin for the transport of phosphate. Here, the expression, purification and crystallization of the phosphate-binding protein PhoX and X-ray data collection at 3.0 Å resolution are described. Biochemical, biophysical and structural data for this protein will be helpful in the elucidation of its function in phosphate uptake and the physiology of the bacterium.

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Purification, crystallization, and preliminary X-ray diffraction analysis of rat kidney annexin V, a calcium-dependent phospholipid-binding protein.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)39600-0 ◽

1990 ◽

Vol 265 (8) ◽

pp. 4567-4569

Author(s):

B A Seaton ◽

J F Head ◽

M A Kaetzel ◽

J R Dedman

Keyword(s):

Diffraction Analysis ◽

Binding Protein ◽

Rat Kidney ◽

Annexin V ◽

X Ray Diffraction ◽

X Ray ◽

Phospholipid Binding ◽

Calcium Dependent

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Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the VP8* carbohydrate-binding protein of the human rotavirus strain Wa

Acta Crystallographica Section F Structural Biology and Crystallization Communications ◽

10.1107/s1744309105032999 ◽

2005 ◽

Vol 61 (11) ◽

pp. 989-993 ◽

Cited By ~ 8

Author(s):

Mark J. Kraschnefski ◽

Stacy A. Scott ◽

Gavan Holloway ◽

Barbara S. Coulson ◽

Mark von Itzstein ◽

...

Keyword(s):

Diffraction Analysis ◽

Binding Protein ◽

Carbohydrate Binding ◽

X Ray Diffraction ◽

X Ray ◽

Carbohydrate Binding Protein ◽

Human Rotavirus

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Expression, purification, crystallization and preliminary X-ray diffraction analysis of the novel modular DNA-binding protein BurrH in its apo form and in complex with its target DNA

Acta Crystallographica Section F Structural Biology Communications ◽

10.1107/s2053230x13033037 ◽

2013 ◽

Vol 70 (1) ◽

pp. 87-91 ◽

Cited By ~ 9

Author(s):

Stefano Stella ◽

Rafael Molina ◽

Claudia Bertonatti ◽

Alexandre Juillerrat ◽

Guillermo Montoya

Keyword(s):

Dna Binding ◽

Diffraction Analysis ◽

Binding Protein ◽

Dna Binding Protein ◽

The Novel ◽

X Ray Diffraction ◽

X Ray ◽

Target Dna

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Crystallization and preliminary X-ray diffraction analysis of human calcium-binding protein S100A12

Acta Crystallographica Section D Biological Crystallography ◽

10.1107/s0907444999014936 ◽

2000 ◽

Vol 56 (2) ◽

pp. 189-191 ◽

Cited By ~ 8

Author(s):

Olga V. Moroz ◽

Alfred A. Antson ◽

G. Guy Dodson ◽

Keith S. Wilson ◽

Inge Skibshøj ◽

...

Keyword(s):

Diffraction Analysis ◽

Calcium Binding ◽

Binding Protein ◽

Calcium Binding Protein ◽

X Ray Diffraction ◽

X Ray

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Phosphate-binding protein fromPolaromonasJS666: purification, characterization, crystallization and sulfur SAD phasing

Acta Crystallographica Section F Structural Biology Communications ◽

10.1107/s2053230x17007373 ◽

2017 ◽

Vol 73 (6) ◽

pp. 342-346 ◽

Cited By ~ 1

Author(s):

Vanessa R. Pegos ◽

Louis Hey ◽

Jacob LaMirande ◽

Rachel Pfeffer ◽

Rosalie Lipsh ◽

...

Keyword(s):

Binding Mode ◽

Binding Activity ◽

X Ray Diffraction ◽

Phosphate Binding ◽

X Ray ◽

Sad Phasing ◽

Binding Residues ◽

Phosphate Binding Protein ◽

Binding Cleft ◽

Aspartate Residue

Phosphate-binding proteins (PBPs) are key proteins that belong to the bacterial ABC-type phosphate transporters. PBPs are periplasmic (or membrane-anchored) proteins that capture phosphate anions from the environment and release them to the transmembrane transporter. Recent work has suggested that PBPs have evolved for high affinity as well as high selectivity. In particular, a short, unique hydrogen bond between the phosphate anion and an aspartate residue has been shown to be critical for selectivity, yet is not strictly conserved in PBPs. Here, the PBP fromPolaromonasJS666 is focused on. Interestingly, this PBP is predicted to harbor different phosphate-binding residues to currently known PBPs. Here, it is shown that the PBP fromPolaromonasJS666 is capable of binding phosphate, with a maximal binding activity at pH 8. Its structure is expected to reveal its binding-cleft configuration as well as its phosphate-binding mode. Here, the expression, purification, characterization, crystallization and X-ray diffraction data collection to 1.35 Å resolution of the PBP fromPolaromonasJS666 are reported.

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