Faculty Opinions recommendation of Helicobacter pylori UreE, a urease accessory protein: specific Ni(2+)- and Zn(2+)-binding properties and interaction with its cognate UreG.

2009 ◽  
Author(s):  
Michael Maroney
Keyword(s):  
Helicobacter Pylori ◽  
Accessory Protein ◽  
Binding Properties ◽  
Urease Accessory Protein

scholarly journals Nickel binding properties of Helicobacter pylori UreF, an accessory protein in the nickel-based activation of urease

2013 ◽  
Vol 19 (3) ◽  
pp. 319-334 ◽  
Author(s):  
Barbara Zambelli ◽  
Andrea Berardi ◽  
Vlad Martin-Diaconescu ◽  
Luca Mazzei ◽  
Francesco Musiani ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Accessory Protein ◽  
Binding Properties ◽  
Nickel Binding

scholarly journals Crystal structure of a truncated urease accessory protein UreF from Helicobacter pylori

2010 ◽  
Vol 78 (13) ◽  
pp. 2839-2848 ◽  
Author(s):  
Robert Lam ◽  
Vladimir Romanov ◽  
Kathy Johns ◽  
Kevin P. Battaile ◽  
Jean Wu-Brown ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Helicobacter Pylori ◽  
Accessory Protein ◽  
Urease Accessory Protein

scholarly journals Helicobacter pylori UreE, a urease accessory protein: specific Ni2+- and Zn2+-binding properties and interaction with its cognate UreG

2009 ◽  
Vol 422 (1) ◽  
pp. 91-100 ◽  
Author(s):  
Matteo Bellucci ◽  
Barbara Zambelli ◽  
Francesco Musiani ◽  
Paola Turano ◽  
Stefano Ciurli
Keyword(s):  
Helicobacter Pylori ◽  
Light Scattering ◽  
Metal Ion ◽  
Site Directed Mutagenesis ◽  
Size Exclusion ◽  
Titration Calorimetry ◽  
Directed Mutagenesis ◽  
Accessory Protein ◽  
Oligomeric State ◽  
Binding Properties

The persistence of Helicobacter pylori in the hostile environment of the human stomach is ensured by the activity of urease. The essentiality of Ni2+ for this enzyme demands proper intracellular trafficking of this metal ion. The metallo-chaperone UreE promotes Ni2+ insertion into the apo-enzyme in the last step of urease maturation while facilitating concomitant GTP hydrolysis. The present study focuses on the metal-binding properties of HpUreE (Helicobacter pylori UreE) and its interaction with the related accessory protein HpUreG, a GTPase involved in the assembly of the urease active site. ITC (isothermal titration calorimetry) showed that HpUreE binds one equivalent of Ni2+ (Kd=0.15 μM) or Zn2+ (Kd=0.49 μM) per dimer, without modification of the protein oligomeric state, as indicated by light scattering. Different ligand environments for Zn2+ and Ni2+, which involve crucial histidine residues, were revealed by site-directed mutagenesis, suggesting a mechanism for discriminating metal-ion-specific binding. The formation of a HpUreE–HpUreG protein complex was revealed by NMR spectroscopy, and the thermodynamics of this interaction were established using ITC. A role for Zn2+, and not for Ni2+, in the stabilization of this complex was demonstrated using size-exclusion chromatography, light scattering, and ITC experiments. A calculated viable structure for the complex suggested the presence of a novel binding site for Zn2+, actually detected using ITC and site-directed mutagenesis. The results are discussed in relation to available evidence of a UreE–UreG functional interaction in vivo. A possible role for Zn2+ in the Ni2+-dependent urease system is envisaged.

scholarly journals A Comparative Study on Nickel Binding to Hpn-like Polypeptides from Two Helicobacter pylori Strains

2021 ◽  
Vol 22 (24) ◽  
pp. 13210
Author(s):  
Danuta Witkowska ◽  
Agnieszka Szebesczyk ◽  
Joanna Wątły ◽  
Michał Braczkowski ◽  
Magdalena Rowińska-Żyrek
Keyword(s):  
Helicobacter Pylori ◽  
Comparative Study ◽  
Potentiometric Titration ◽  
Isothermal Titration Calorimetry ◽  
Titration Calorimetry ◽  
Binding Properties ◽  
Protein Fragments ◽  
Nickel Binding ◽  
Independent Reaction

Combined potentiometric titration and isothermal titration calorimetry (ITC) methods were used to study the interactions of nickel(II) ions with the N-terminal fragments and histidine-rich fragments of Hpn-like protein from two Helicobacter pylori strains (11637 and 26695). The ITC measurements were performed at various temperatures and buffers in order to extract proton-independent reaction enthalpies of nickel binding to each of the studied protein fragments. We bring up the problem of ITC results of nickel binding to the Hpn-like protein being not always compatible with those from potentiometry and MS regarding the stoichiometry and affinity. The roles of the ATCUN motif and multiple His and Gln residues in Ni(II) binding are discussed. The results provided the possibility to compare the Ni(II) binding properties between N-terminal and histidine-rich part of Hpn-like protein and between N-terminal parts of two Hpn-like strains, which differ mainly in the number of glutamine residues.

scholarly journals Biochemical and genetic analyses of the oomycetePythium insidiosumprovide new insights into clinical identification and urease-based evolution of metabolism-related traits

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e4821 ◽  
Author(s):  
Theerapong Krajaejun ◽  
Thidarat Rujirawat ◽  
Teerat Kanpanleuk ◽  
Pitak Santanirand ◽  
Tassanee Lohnoo ◽  
...  
Keyword(s):  
Initial Assessment ◽  
Surgical Removal ◽  
Accessory Protein ◽  
Pythium Insidiosum ◽  
Biochemical Characteristics ◽  
Biochemical Assays ◽  
Protein Encoding ◽  
Encoding Genes ◽  
Hydrolysis Of ◽  
Urease Accessory Protein

The oomycete microorganism,Pythium insidiosum, causes the life-threatening infectious condition, pythiosis, in humans and animals worldwide. Affected individuals typically endure surgical removal of the infected organ(s). Detection ofP. insidiosumby the established microbiological, immunological, or molecular methods is not feasible in non-reference laboratories, resulting in delayed diagnosis. Biochemical assays have been used to characterizeP. insidiosum, some of which could aid in the clinical identification of this organism. Although hydrolysis of maltose and sucrose has been proposed as the key biochemical feature useful in discriminatingP. insidiosumfrom other oomycetes and fungi, this technique requires a more rigorous evaluation involving a wider selection ofP. insidiosumstrains. Here, we evaluated 10 routinely available biochemical assays for characterization of 26P. insidiosumstrains, isolated from different hosts and geographic origins. Initial assessment revealed diverse biochemical characteristics across theP. insidiosumstrains tested. Failure to hydrolyze sugars is observed, especially in slow-growing strains. Because hydrolysis of maltose and sucrose varied among different strains, use of the biochemical assays for identification ofP. insidiosumshould be cautioned. The ability ofP. insidiosumto hydrolyze urea is our focus, because this metabolic process relies on the enzyme urease, an important virulence factor of other pathogens. The ability to hydrolyze urea varied amongP. insidiosumstrains and was not associated with growth rates. Genome analyses demonstrated that urease- and urease accessory protein-encoding genes are present in both urea-hydrolyzing and non-urea-hydrolyzing strains ofP. insidiosum. Urease genes are phylogenetically conserved inP. insidiosumand related oomycetes, while the presence of urease accessory protein-encoding genes is markedly diverse in these organisms. In summary, we dissected biochemical characteristics and drew new insights into clinical identification and urease-related evolution ofP. insidiosum.

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