Cloning, characterisation and crystallisation of a diadenosine 5′,5‴-P1,P4-tetraphosphate pyrophosphohydrolase from Caenorhabditis elegans

Abdelghany, Hend M.; Gasmi, Lakhdar; Cartwright, Jared; Bailey, Scott; Rafferty, John B.; McLennan, Alexander G.

doi:10.1016/s0167-4838(01)00263-1

Cited by 32 publications

(48 citation statements)

References 35 publications

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“…Ap 4 A Hydrolase Assay-Ap 4 A hydrolase activity was measured using a luciferase-based bioluminescence assay as previously described (6). One ng enzyme protein was used in each case, except for K83M (10 ng), K79M (20 ng), E52Q and E103Q (60 ng), and E56Q (600 ng).…”

Section: Cloning Of C Elegans Ap 4 a Hydrolase As A Glutathione S-trmentioning

confidence: 99%

“…Some are highly specific whereas others appear to have a broad substrate range in vitro (3)(4)(5). The Nudix Ap 4 A hydrolases can be further subdivided into "plant" and "animal"-types, according to their primary structure (6). The plant-type includes enzymes from the Proteobacteria that have in some cases been shown to be associated with the invasion of mammalian cells, whereas the animal-type includes putative Ap 4 A hydrolases from Archaea (6 -10).…”

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confidence: 99%

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Analysis of the Catalytic and Binding Residues of the Diadenosine Tetraphosphate Pyrophosphohydrolase from Caenorhabditis elegans by Site-directed Mutagenesis

Abdelghany¹,

Bailey²,

Blackburn³

et al. 2003

Journal of Biological Chemistry

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Section: Cloning Of C Elegans Ap 4 a Hydrolase As A Glutathione S-trmentioning

confidence: 99%

mentioning

confidence: 99%

Analysis of the Catalytic and Binding Residues of the Diadenosine Tetraphosphate Pyrophosphohydrolase from Caenorhabditis elegans by Site-directed Mutagenesis

Abdelghany¹,

Bailey²,

Blackburn³

et al. 2003

Journal of Biological Chemistry

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“…Homogeneous recombinant (asymmetrical) Ap 4 A hydrolases (EC 3.6.1.17) from human [24], narrow-leafed lupin (Lupinus angustifolius) [25] and Caenorhabditis elegans [26] were obtained as described. The lupin enzyme was kindly donated by Dr D. Maksel and Dr K. Gayler (University of Melbourne, Melbourne, Australia).…”

Section: Experimental Enzymesmentioning

confidence: 99%

“…Asymmetrically acting Ap 4 A hydrolases were assayed in an incubation mixture (0.05 ml total volume) containing 50 mM Hepes/KOH (pH 7.6), 0.02 mM dithiothreitol, 5 mM MgCl 2 , 0.05 mM The recently established three-dimensional structures of the lupin Ap 4 A hydrolase [20] and the hydrolase from C. elegans [29], which is closely related to the human enzyme [26], show a number of structural differences within the substrate-binding sites of the plant and animal enzymes. Therefore it is not surprising that recognition of other potential ligands (inhibitors) by these hydrolases differs too.…”

Section: Enzyme Assaysmentioning

confidence: 99%

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Adenosine-5'-O-phosphorylated and adenosine-5'-O-phosphorothioylated polyols as strong inhibitors of (symmetrical) and (asymmetrical) dinucleoside tetraphosphatases

Guranowski¹,

Starzyńska²,

McLennan

et al. 2003

Biochemical Journal

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Dinucleoside 5 ,5 -P 1 ,P n -polyphosphates, and particularly the diadenosine compounds, have been implicated in extracellular purinergic signalling and in various intracellular processes, including DNA metabolism, tumour suppression and stress responses. If permitted to accumulate, they may also be toxic. One approach to understanding their function is through the various specific degradative enzymes that regulate their levels. Eight adenosine-5 -O-phosphorylated polyols (derivatives of glycerol, erythritol and pentaerythritol) and 11 adenosine-5 -O-phosphorothioylated polyols (derivatives of glycerol, erythritol, pentaerythritol, butanediol and pentanediol) have been tested as inhibitors of specific diadenosine tetraphosphate (Ap 4 A) hydrolases. Of these two groups of novel nucleotides, the adenosine-5 -O-phosphorothioylated polyols were generally stronger inhibitors than their adenosine-5 -O-phosphorylated counterparts. 1,4-Di(adenosine-5 -O-phosphorothio) erythritol appeared to be the strongest inhibitor of (asymmetrical) Ap 4 A hydrolases (EC 3.6.1.17) from both lupin and human, with K i values of 0.15 µM and 1.5 µM respectively. Of eight adenosine-5 -Ophosphorylated polyols, 1,4-di(adenosine-5 -O-phospho) erythritol was the only compound that inhibited the lupin enzyme. Two derivatives of pentaerythritol, di(adenosine-5 -O-phosphorothio)-di(phosphorothio) pentaerythritol and tri(adenosine-5 -Ophosphorothio)-phosphorothio-pentaerythritol, proved to be the strongest inhibitors of the prokaryotic (symmetrical) Ap 4 A hydrolase (EC 3.6.1.41) so far reported. The estimated K i values were 0.04 µM and 0.08 µM respectively. All of these inhibitors were competitive with respect to Ap 4 A. These new selectively acting Ap 4 A analogues should prove to be valuable tools for further studies of Ap 4 A function and of the enzymes involved in its metabolism.

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The evolution of function within the Nudix homology clan

Srouji

Park

et al. 2017

Proteins

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The Nudix homology clan encompasses over 80,000 protein domains from all three domains of life, defined by homology to each other. Proteins with a domain from this clan fall into four general functional classes: pyrophosphohydrolases, isopentenyl diphosphate isomerases (IDIs), adenine/guanine mismatch-specific adenine glycosylases (A/G-specific adenine glycosylases), and non-enzymatic activities such as protein/protein interaction and transcriptional regulation. The largest group, pyrophosphohydrolases, encompasses more than 100 distinct hydrolase specificities. To understand the evolution of this vast number of activities, we assembled and analyzed experimental and structural data for 205 Nudix proteins collected from the literature. We corrected erroneous functions or provided more appropriate descriptions for 53 annotations described in the Gene Ontology Annotation database in this family, and propose 275 new experimentally-based annotations. We manually constructed a structure-guided sequence alignment of 78 Nudix proteins. Using the structural alignment as a seed, we then made an alignment of 347 “select” Nudix homology domains, curated from structurally determined, functionally characterized, or phylogenetically important Nudix domains. Based on our review of Nudix pyrophosphohydrolase structures and specificities, we further analyzed a loop region downstream of the Nudix hydrolase motif previously shown to contact the substrate molecule and possess known functional motifs. This loop region provides a potential structural basis for the functional radiation and evolution of substrate specificity within the hydrolase family. Finally, phylogenetic analyses of the 347 select protein domains and of the complete Nudix homology clan revealed general monophyly with regard to function and a few instances of probable homoplasy.

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Cloning, characterisation and crystallisation of a diadenosine 5′,5‴-P1,P4-tetraphosphate pyrophosphohydrolase from Caenorhabditis elegans

Cited by 32 publications

References 35 publications

Analysis of the Catalytic and Binding Residues of the Diadenosine Tetraphosphate Pyrophosphohydrolase from Caenorhabditis elegans by Site-directed Mutagenesis

Analysis of the Catalytic and Binding Residues of the Diadenosine Tetraphosphate Pyrophosphohydrolase from Caenorhabditis elegans by Site-directed Mutagenesis

Adenosine-5'-O-phosphorylated and adenosine-5'-O-phosphorothioylated polyols as strong inhibitors of (symmetrical) and (asymmetrical) dinucleoside tetraphosphatases

The evolution of function within the Nudix homology clan

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