Literature summary for 3.2.1.143 extracted from

Dunstan, M.S.; Barkauskaite, E.; Lafite, P.; Knezevic, C.E.; Brassington, A.; Ahel, M.; Hergenrother, P.J.; Leys, D.; Ahel, I.
Structure and mechanism of a canonical poly(ADP-ribose) glycohydrolase (2012), Nat. Commun., 3, 878.

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Application

Application	Comment	Organism
drug development	the protozoan enzyme represents a good model for human PARG and is therefore likely to prove useful in guiding structure-based discovery of new classes of PARG inhibitors	Tetrahymena thermophila

Cloned(Commentary)

Cloned (Comment)	Organism
phylogenetic analysis, expression of wild-type and selenomethionin-labeled enzymes in Escherichia coli strain Rosetta (DE3)	Tetrahymena thermophila

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged enzyme in complex with ADP-ribose or inhibitor RBPI-3, and as selenomethionine-labeled enzyme, sitting drop vapour diffusion method, 15 mg/ml protein is mixed with a 1:1 molar ratio with ADP-ribose, and with an equal volume of a solution containing 0.15 M potassium thiocyanate, 0.1 M Tris, pH 8.5, and 15% PEG 6000, or with 0.2 M potassium bromide, 0.1 M Tris, pH 7.5, and 15% PEG 4000, X-ray diffraction structure determination and analysis at 1.95 A resolution	Tetrahymena thermophila

Crystallization (Comment)

Organism

purified recombinant His-tagged enzyme in complex with ADP-ribose or inhibitor RBPI-3, and as selenomethionine-labeled enzyme, sitting drop vapour diffusion method, 15 mg/ml protein is mixed with a 1:1 molar ratio with ADP-ribose, and with an equal volume of a solution containing 0.15 M potassium thiocyanate, 0.1 M Tris, pH 8.5, and 15% PEG 6000, or with 0.2 M potassium bromide, 0.1 M Tris, pH 7.5, and 15% PEG 4000, X-ray diffraction structure determination and analysis at 1.95 A resolution

Tetrahymena thermophila

Inhibitors

Inhibitors	Comment	Organism	Structure
RBPI-3	a rhodanine-containing mammalian PARG inhibitor, enzyme-inhibitor structure analysis, overview. RBPI-3 binds predominantly via a pi-pi stacking interaction with Tyr296 and the conserved Phe398. To accommodate the binding of RBPI-3, Phe398 moves into the adenosine binding pocket. The RBPI-3 carboxyl moiety occupies a region corresponding to the ADP-ribose alpha-phosphate group and H-bonds to main chain atoms of Lys365 and Gln254. The RBPI-3 di-chlorobenzyl moiety extends into the solvent and is disordered	Tetrahymena thermophila

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
poly(ADP-D-ribose)n + H2O	Tetrahymena thermophila	the activity is dependent on the conserved glutamate residues	poly(ADP-D-ribose)n-1 + ADP-ribose	-	?

Organism

Organism	UniProt	Comment	Textmining
Tetrahymena thermophila	I6L8L8	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type and selenomethionine-labeled enzymes from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography and gel filtration	Tetrahymena thermophila

Reaction

Reaction	Comment	Organism	Reaction ID
(ADP-ribose)n + H2O = (ADP-ribose)n-1 + ADP-ribose	canonical enzyme catalytic mechanism, modelling, overview	Tetrahymena thermophila	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
poly(ADP-D-ribose)n + H2O	the activity is dependent on the conserved glutamate residues	Tetrahymena thermophila	poly(ADP-D-ribose)n-1 + ADP-ribose	-	?

Subunits

Subunits	Comment	Organism
More	the enzyme structure consists of a macrodomain sandwiched between a large N-terminal accessory domain and a smaller carboxy-terminal extension, structure overview	Tetrahymena thermophila

Synonyms

Synonyms	Comment	Organism
PAR glycohydrolase	-	Tetrahymena thermophila
PARG	-	Tetrahymena thermophila

General Information

General Information	Comment	Organism
evolution	canonical poly(ADP-ribose) glycohydrolase is a highly conserved protein found in organisms ranging from protozoa to humans, phylogenetic analysis. The full-length enzyme from Tetrahymena thermophila is highly similar to the minimal catalytic region of thhe human enzyme, but it lacks the obvious RS/MTS motif	Tetrahymena thermophila
additional information	enzyme structure overview	Tetrahymena thermophila
physiological function	the reversion of poly(ADP-ribosyl)ation is catalysed by poly(ADP-ribose) glycohydrolase, which specifically targets the unique PAR (1''-2') ribose-ribose bonds	Tetrahymena thermophila