Contribution of the two domains of <i>E. coli</i> 5′‐nucleotidase to substrate specificity and catalysis

Krug, Ulrike; Patzschke, Rica; Zebisch, M.; Balbach, Jochen; Sträter, Norbert

doi:10.1016/j.febslet.2013.01.010

Cited by 14 publications

(16 citation statements)

References 23 publications

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“…The main function of UshA was therefore proposed to be as a metabolic enzyme. Thereafter, it was demonstrated that NTP, NDP, NMP, and nucleotide sugars were also UshA substrates, but the biological function of this enzyme remains unclear (39,42).…”

Section: Discussionmentioning

confidence: 99%

Extracellular Nucleotide Catabolism by the Group B Streptococcus Ectonucleotidase NudP Increases Bacterial Survival in Blood

Firon

Dinis

Raynal

et al. 2014

Journal of Biological Chemistry

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Background: Ectonucleotidases regulate extracellular nucleotide concentration. Results: The NudP ecto-5Ј-nucleotidase of Streptococcus agalactiae has specific substrate specificities necessary for survival in blood and organ colonization. Conclusion: Extracellular nucleotide catabolism is involved in the control of Group B streptococcal pathogenesis. Significance: Bacterial pathogens exploit different enzymatic specificities to subvert extracellular nucleotide signaling.

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Section: Discussionmentioning

confidence: 99%

Extracellular Nucleotide Catabolism by the Group B Streptococcus Ectonucleotidase NudP Increases Bacterial Survival in Blood

Firon

Dinis

Raynal

et al. 2014

Journal of Biological Chemistry

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“…The experimental procedure is detailed in the supplement. Briefly, after expression in E. coli and chromatographic purification (Schultz-Heienbrok et al, 2004; Krug et al, 2013), a tenfold excess of MTSSL over protein was added in two steps with incubation at room temperature and 4°C. Unreacted spin label was removed by a desalting column.…”

Section: Methodsmentioning

confidence: 99%

“…It is a two-domain enzyme with a dimetal center located in the N-terminal domain where various substrates including nucleotides, nucleotide sugars or dinucleotides are hydrolyzed (Glaser et al, 1967; Neu, 1967; Ruiz et al, 1989; Knöfel and Sträter, 1999, 2001b; Schultz-Heienbrok et al, 2005; Krug et al, 2013). Substrates bind to the C-terminal domain in the open state and a 96° rotation of this domain around its center enables hydrolysis via formation of the closed state (Figure 1).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the Domain Orientations of E. coli 5′-Nucleotidase by Fitting an Ensemble of Conformers to DEER Distance Distributions

et al. 2016

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Summary Escherichia coli 5’-nucleotidase is a two-domain enzyme exhibiting a unique 96° domain motion that is required for catalysis. Here we present an integrated structural biology study that combines DEER distance distributions with structural information from X-ray crystallography and computational biology to describe the population of presumably almost isoenergetic open and closed states in solution. Ensembles of models which represent the experimental distance distributions best are determined by a Monte Carlo search algorithm. As a result, predominantly open conformations are observed in the unliganded state indicating that the majority of enzyme molecules await substrate binding for the catalytic cycle. The addition of a substrate analog yields ensembles with an almost equal mixture of open and closed states. Thus, in the presence of substrate efficient catalysis is provided by the simultaneous appearance of open conformers (binding substrate or releasing product) and closed conformers (enabling the turnover of the substrate).

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“…These two domains (the metallophosphoesterase and 5′-nucleotidase domains) are linked by an α-helix. The molecular structures and catalytic mechanisms of enzymes of this type were thoroughly investigated with Escherichia coli UshA used as a representative (Knofel and Strater 2001 ; Krug et al 2013 ). The C-terminal domain is responsible for binding the substrate base, while the N-terminal domain is responsible for the core catalytic steps.…”

Section: Classification Of Microbial 5′-nucleotidases Their Molecular Structures and Preferable Substratesmentioning

confidence: 99%

Microbial 5′-nucleotidases: their characteristics, roles in cellular metabolism, and possible practical applications

Zakataeva

2021

Appl Microbiol Biotechnol

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Abstract5′-Nucleotidases (EC 3.1.3.5) are enzymes that catalyze the hydrolytic dephosphorylation of 5′-ribonucleotides and 5′-deoxyribonucleotides to their respective nucleosides and phosphate. Most 5′-nucleotidases have broad substrate specificity and are multifunctional enzymes capable of cleaving phosphorus from not only mononucleotide phosphate molecules but also a variety of other phosphorylated metabolites. 5′-Nucleotidases are widely distributed throughout all kingdoms of life and found in different cellular locations. The well-studied vertebrate 5′-nucleotidases play an important role in cellular metabolism. These enzymes are involved in purine and pyrimidine salvage pathways, nucleic acid repair, cell-to-cell communication, signal transduction, control of the ribo- and deoxyribonucleotide pools, etc. Although the first evidence of microbial 5′-nucleotidases was obtained almost 60 years ago, active studies of genetic control and the functions of microbial 5′-nucleotidases started relatively recently. The present review summarizes the current knowledge about microbial 5′-nucleotidases with a focus on their diversity, cellular localizations, molecular structures, mechanisms of catalysis, physiological roles, and activity regulation and approaches to identify new 5′-nucleotidases. The possible applications of these enzymes in biotechnology are also discussed.Key points• Microbial 5′-nucleotidases differ in molecular structure, hydrolytic mechanism, and cellular localization.• 5′-Nucleotidases play important and multifaceted roles in microbial cells.• Microbial 5′-nucleotidases have wide range of practical applications. Graphical abstract

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Contribution of the two domains of E. coli 5′‐nucleotidase to substrate specificity and catalysis

Cited by 14 publications

References 23 publications

Extracellular Nucleotide Catabolism by the Group B Streptococcus Ectonucleotidase NudP Increases Bacterial Survival in Blood

Extracellular Nucleotide Catabolism by the Group B Streptococcus Ectonucleotidase NudP Increases Bacterial Survival in Blood

Characterization of the Domain Orientations of E. coli 5′-Nucleotidase by Fitting an Ensemble of Conformers to DEER Distance Distributions

Microbial 5′-nucleotidases: their characteristics, roles in cellular metabolism, and possible practical applications

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