Abstract
Regulation of cellular levels of ADP-ribose is important in preventing nonenzymatic ADP-ribosylation of proteins. The Escherichia coli ADP-ribose pyrophosphatase, a Nudix enzyme, catalyzes the hydrolysis of ADP-ribose to ribose-5-P and AMP, compounds that can be recycled as part of nucleotide metabolism. The structures of the apo enzyme, the active enzyme and the complex with ADP-ribose were determined to 1.9 A, 2.7 A and 2.3 A, respectively. The structures reveal a symmetric homodimer with two equivalent catalytic sites, each formed by residues of both monomers, requiring dimerization through domain swapping for substrate recognition and catalytic activity. The structures also suggest a role for the residues conserved in each Nudix subfamily. The Nudix motif residues, folded as a loop-helix-loop tailored for pyrophosphate hydrolysis, compose the catalytic center; residues conferring substrate specificity occur in regions of the sequence removed from the Nudix motif. This segregation of catalytic and recognition roles provides versatility to the Nudix family.
Publication types
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adenosine Diphosphate Ribose / metabolism
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Amino Acid Motifs
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Amino Acid Sequence
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Apoenzymes / chemistry
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Apoenzymes / metabolism
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Bacterial Proteins / chemistry
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Bacterial Proteins / metabolism
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Binding Sites
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Cations, Divalent / metabolism
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Conserved Sequence
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Crystallography, X-Ray
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Dimerization
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Enzyme Activation
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Escherichia coli / enzymology*
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Escherichia coli Proteins*
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Hydrolysis
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Magnesium / metabolism
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Models, Molecular
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Molecular Sequence Data
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Nudix Hydrolases
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Phosphoric Monoester Hydrolases / chemistry
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Phosphoric Monoester Hydrolases / metabolism
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Protein Structure, Tertiary
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Pyrophosphatases / chemistry*
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Pyrophosphatases / metabolism*
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Sequence Alignment
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Structure-Activity Relationship
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Substrate Specificity
Substances
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Apoenzymes
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Bacterial Proteins
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Cations, Divalent
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Escherichia coli Proteins
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Adenosine Diphosphate Ribose
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Phosphoric Monoester Hydrolases
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Pyrophosphatases
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mutT protein, E coli
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ADPribose pyrophosphatase
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Magnesium
Associated data
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PDB/1G0S
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PDB/1G9Q
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PDB/1GA7