Crystallization and preliminary X-ray diffraction analysis of a high-affinity phosphate-binding protein endowed with phosphatase activity fromPseudomonas aeruginosaPAO1

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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The high-affinity phosphate-binding protein PstS is accumulated under high fructose concentrations and mutation of the corresponding gene affects differentiation in Streptomyces lividans

Microbiology ◽

10.1099/mic.0.27983-0 ◽

2005 ◽

Vol 151 (8) ◽

pp. 2583-2592 ◽

Cited By ~ 42

Author(s):

Margarita Díaz ◽

Ana Esteban ◽

José Manuel Fernández-Abalos ◽

Ramón I. Santamaría

Keyword(s):

Culture Media ◽

Binding Protein ◽

Protein Pattern ◽

Phosphate Transport ◽

Streptomyces Lividans ◽

Null Mutant ◽

Phosphate Binding ◽

Genetic Switch ◽

High Affinity ◽

Phosphate Binding Protein

The secreted protein pattern of Streptomyces lividans depends on the carbon source present in the culture media. One protein that shows the most dramatic change is the high-affinity phosphate-binding protein PstS, which is strongly accumulated in the supernatant of liquid cultures containing high concentrations (>3 %) of certain sugars, such as fructose, galactose and mannose. The promoter region of this gene and that of its Streptomyces coelicolor homologue were used to drive the expression of a xylanase in S. lividans that was accumulated in the culture supernatant when grown in the presence of fructose. PstS accumulation was dramatically increased in a S. lividans polyphosphate kinase null mutant (Δppk) and was impaired in a deletion mutant lacking phoP, the transcriptional regulator gene of the two-component phoR-phoP system that controls the Pho regulon. Deletion of the pstS genes in S. lividans and S. coelicolor impaired phosphate transport and accelerated differentiation and sporulation on solid media. Complementation with a single copy in a S. lividans pstS null mutant returned phosphate transport and sporulation to levels similar to those of the wild-type strain. The present work demonstrates that carbon and phosphate metabolism are linked in the regulation of genes and that this can trigger the genetic switch towards morphogenesis.

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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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High-affinity phosphate-binding protein (PBP) for phosphorous recovery: proof of concept using recombinantEscherichia coli

FEMS Microbiology Letters ◽

10.1093/femsle/fnw240 ◽

2016 ◽

Vol 363 (20) ◽

pp. fnw240 ◽

Cited By ~ 5

Author(s):

Yu Yang ◽

Wendy Ballent ◽

Brooke K. Mayer

Keyword(s):

Binding Protein ◽

Proof Of Concept ◽

Phosphate Binding ◽

High Affinity ◽

Phosphate Binding Protein

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Crystallization and preliminary X-ray diffraction analysis of a DING protein fromPseudomonas aeruginosaPA14

Acta Crystallographica Section F Structural Biology and Crystallization Communications ◽

10.1107/s1744309113005356 ◽

2013 ◽

Vol 69 (4) ◽

pp. 425-429 ◽

Cited By ~ 2

Author(s):

Ahmed Djeghader ◽

Guillaume Gotthard ◽

Andrew Suh ◽

Daniel Gonzalez ◽

Ken Scott ◽

...

Keyword(s):

Biochemical Characterization ◽

Protein Structures ◽

Biological Processes ◽

X Ray Diffraction ◽

Phosphate Binding ◽

Structural Determinants ◽

X Ray ◽

High Affinity ◽

Ding Protein ◽

Hiv 1

DING proteins form an emergent family of proteins consisting of an increasing number of homologues that have been identified in all kingdoms of life. They belong to the superfamily of phosphate-binding proteins and exhibit a high affinity for phosphate. In eukaryotes, DING proteins have been isolated by virtue of their implication in several diseases and biological processes. Some of them are potent inhibitors of HIV-1 replication/transcription, raising the question of their potential involvement in the human defence system. Recently, a protein fromPseudomonas aeruginosastrain PA14, named PA14DING or LapC, belonging to the DING family has been identified. The structure of PA14DING, combined with detailed biochemical characterization and comparative analysis with available DING protein structures, will be helpful in understanding the structural determinants implicated in the inhibition of HIV-1 by DING proteins. Here, the expression, purification and crystallization of PA14DING and the collection of X-ray data to 1.9 Å resolution are reported.

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