Characterization of the
            <i>Klebsiella aerogenes</i>
            Urease Accessory Protein UreD in Fusion with the Maltose Binding Protein

Carter, Eric L.; Hausinger, Robert P.

doi:10.1128/jb.01426-09

Cited by 32 publications

(87 citation statements)

References 34 publications

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“…Due to the insolubility of UreD when it is overexpressed alone, there is no reported crystal structure of UreD and it is the least characterized of the urease accessory proteins. However, recently, investigators have found a maltose binding protein-UreD fusion to be soluble and useful in characterization studies (4). Consistent with previous reports, these investigators found UreD to bind both UreF and UreG in vivo.…”

Section: Discussionsupporting

confidence: 77%

“…The total protein concentration in the lysates was determined using the Bio-Rad DC protein assay, according to the manufacturer's specifications and with bovine serum albumin as the standard. A lysate volume containing 50 g protein was combined with the phenol-hypochlorite assay buffer, consisting of 50 mM HEPES, pH 7.5, and 25 mM urea, and the mixture was compared to standards prepared from NH 4 Cl. E. coli MG1655 was used as a negative control, and EHEC O157:H7 strain IN1 and/or Mo28 was used as a positive control.…”

Section: Methodsmentioning

confidence: 99%

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Functional and Phylogenetic Analysis of ureD in Shiga Toxin-Producing Escherichia coli

2011

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Enterohemorrhagic Escherichia coli (EHEC) is a food-borne pathogen that can cause severe health complications and utilizes a much lower infectious dose than other E. coli pathotypes. Despite having an intact ure locus, ureDABCEFG, the majority of EHEC strains are phenotypically urease negative under tested conditions. Urease activity potentially assists with survival fitness by enhancing acid tolerance during passage through the stomach or by aiding with colonization in either human or animal reservoirs. Previously, in the EHEC O157:H7 Sakai strain, a point mutation in ureD, encoding a urease chaperone protein, was identified, resulting in a substitution of an amber stop codon for glutamine. This single nucleotide polymorphism (SNP) is observed in the majority of EHEC O157:H7 isolates and correlates with a negative urease phenotype in vitro. We demonstrate that the lack of urease activity in vitro is not solely due to the amber codon in ureD. Our analysis has identified two additional SNPs in ureD affecting amino acid positions 38 and 205, in both cases determining whether the encoded amino acid is leucine or proline. Phylogenetic analysis based on Ure protein sequences from a variety of urease-encoding bacteria demonstrates that the proline at position 38 is highly conserved among Gram-negative bacteria. Experiments reveal that the L38P substitution enhances urease enzyme activity; however, the L205P substitution does not. Multilocus sequence typing analysis for a variety of Shiga toxin-producing E. coli isolates combined with the ureD sequence reveals that except for a subset of the O157:H7 strains, neither the in vitro urease-positive phenotype nor the ureD sequence is phylogenetically restricted.

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Section: Discussionsupporting

confidence: 77%

Section: Methodsmentioning

confidence: 99%

Functional and Phylogenetic Analysis of ureD in Shiga Toxin-Producing Escherichia coli

2011

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“…UreF was reported to form a complex with UreH(D) (9 -12), and the two proteins interact with UreG to form the heterotrimeric complex UreG-UreFUreH(D) (12,13). UreG is a SIMIBI (after signal recognition particle, MinD and BioD) class GTPase and is homologous to HypB, a hydrogenase maturation factor responsible for nickel delivery (14).…”

mentioning

confidence: 99%

“…Urease activation was inhibited by addition of the nonhydrolysable GTP analog, suggesting that GTPase activity of UreG is essential for urease activation (15). The apo-urease can form a complex with UreG-UreF-UreH(D) or its components of UreH(D) and UreF-UreH(D) (12,13,16,17). It has been shown that apo-urease can be activated in vitro by adding an excess amount of both carbon dioxide and nickel ion (18).…”

mentioning

confidence: 99%

Assembly of Preactivation Complex for Urease Maturation in Helicobacter pylori

Fong

Wong

Chuck

et al. 2011

Journal of Biological Chemistry

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Background: Maturation of urease is assisted by urease accessory proteins UreE, UreF, UreG, and UreH. Results: Crystal structure of UreF-UreH complex revealed conformational changes of UreF upon complex formation. Conclusion: Mutagenesis study confirmed that the conformational changes in UreF are essential for recruitment of UreG to the heterotrimeric complex of UreG-UreF-UreH. Significance: Our results provide a structural basis for understanding urease maturation.

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“…To understand the activation mechanisms of ureases and the roles of accessory proteins in the posttranslational modifications of ureases, many different microbial nickel-containing ureases have been identified and characterized. Although the structural subunits and enzyme active sites have been well studied (3)(4)(5)(10)(11)(12)(13)(14)(15), the function of each accessory protein and the urease maturation mechanism are still poorly understood (3,4,(16)(17)(18)(19)(20).…”

mentioning

confidence: 99%

A Bacillus paralicheniformis Iron-Containing Urease Reduces Urea Concentrations in Rice Wine

Liu

Chen

Yuan

et al. 2017

Appl Environ Microbiol

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Urease, a nickel-containing metalloenzyme, was the first enzyme to be crystallized and has a prominent position in the history of biochemistry. In the present study, we identified a nickel urease gene cluster, ureABCEFGDH, in Bacillus paralicheniformis ATCC 9945a and characterized it in Escherichia coli. Enzymatic assays demonstrate that this oxygen-stable urease is also an iron-containing acid urease. Heterologous expression assays of UreH suggest that this accessory protein is involved in the transmembrane transportation of nickel and iron ions. Moreover, this iron-containing acid urease has a potential application in the degradation of urea in rice wine. The present study not only enhances our understanding of the mechanism of activation of urease but also provides insight into the evolution of metalloenzymes.IMPORTANCE An iron-containing, oxygen-stable acid urease from B. paralicheniformis ATCC 9945a with good enzymatic properties was characterized. This acid urease shows activities toward both urea and ethyl carbamate. After digestion with 6 U/ml urease, approximately 92% of the urea in rice wine was removed, suggesting that this urease has great potential in the food industry.

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Characterization of the Klebsiella aerogenes Urease Accessory Protein UreD in Fusion with the Maltose Binding Protein

Cited by 32 publications

References 34 publications

Functional and Phylogenetic Analysis of ureD in Shiga Toxin-Producing Escherichia coli

Functional and Phylogenetic Analysis of ureD in Shiga Toxin-Producing Escherichia coli

Assembly of Preactivation Complex for Urease Maturation in Helicobacter pylori

A Bacillus paralicheniformis Iron-Containing Urease Reduces Urea Concentrations in Rice Wine

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