Literature summary for 3.2.1.143 extracted from

Kim, I.K.; Kiefer, J.R.; Ho, C.M.; Stegeman, R.A.; Classen, S.; Tainer, J.A.; Ellenberger, T.
Structure of mammalian poly(ADP-ribose) glycohydrolase reveals a flexible tyrosine clasp as a substrate-binding element (2012), Nat. Struct. Mol. Biol., 19, 653-656.

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Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of His6-tagged truncated enzyme comprising residues 385-972 in Escherichia coli strain BL21 co-expressing GroESL chaperone	Rattus norvegicus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified truncated native and selenomethionine-labeled enzymes comprising residues 385-972 and lacking the A-domain, free or in complex with adenosine diphosphate (hydroxymethyl) pyrrolidinediol, hanging drop vapour diffusion method, mixing of 15 mg/ml protein in 25 mM HEPES, pH 7.5, 150 mM NaCl, 5% glycerol and 2 mM DTT, with an equal volume of well solution containing 16-20% w/v PEG 2000 monomethylether, 0.1 M Tris-HCl, pH 7.5, 0.1 M NaCl, and 0.2 M potassium thiocyanate, 22°C, X-ray diffraction structure determination and analysis at 1.95 A resolution	Rattus norvegicus

Crystallization (Comment)

Organism

purified truncated native and selenomethionine-labeled enzymes comprising residues 385-972 and lacking the A-domain, free or in complex with adenosine diphosphate (hydroxymethyl) pyrrolidinediol, hanging drop vapour diffusion method, mixing of 15 mg/ml protein in 25 mM HEPES, pH 7.5, 150 mM NaCl, 5% glycerol and 2 mM DTT, with an equal volume of well solution containing 16-20% w/v PEG 2000 monomethylether, 0.1 M Tris-HCl, pH 7.5, 0.1 M NaCl, and 0.2 M potassium thiocyanate, 22°C, X-ray diffraction structure determination and analysis at 1.95 A resolution

Rattus norvegicus

Inhibitors

Inhibitors	Comment	Organism	Structure
adenosine diphosphate (hydroxymethyl) pyrrolidinediol	ADP-HPD, tight binding inhibitor, binding structure, overview	Rattus norvegicus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
additional information	Rattus norvegicus	the enzyme functions as an endo-glycosidase to release oligo(ADP-ribose) and as an exo-glycosidase to release ADP-ribose. Long poly(ADP-ribose) polymers are efficiently hydrolyzed by a combination of endo- and exo-glycosidic activity, whereas smaller digestion products are poor substrates for the enzyme allowing release of oligo(ADP-ribose) chains that are ligands for histones and DNA repair and damage checkpoint proteins such as XRCC1 and p53	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Rattus norvegicus	Q9QYM2	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged truncated enzyme, native and selenomethionine-labeled, from Escherichia coli strain BL21 by nickel affinity and heparin affinity chromatography, and gel filtration	Rattus norvegicus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	the enzyme functions as an endo-glycosidase to release oligo(ADP-ribose) and as an exo-glycosidase to release ADP-ribose. Long poly(ADP-ribose) polymers are efficiently hydrolyzed by a combination of endo- and exo-glycosidic activity, whereas smaller digestion products are poor substrates for the enzyme allowing release of oligo(ADP-ribose) chains that are ligands for histones and DNA repair and damage checkpoint proteins such as XRCC1 and p53	Rattus norvegicus	?	-	?

Subunits

Subunits	Comment	Organism
More	structure analysis and comparisons, overview. The enzyme comprises an N-terminal regulatory and targeting domain (A-domain; residues 1-456), a central mitochondrial targeting sequence (MTS, residues 457-472), and a C-terminal catalytic domain (residues 473-972). The rPARG385 catalytic domain adopts a beanshaped structure with a deep central cleft containing the conserved PARG-signature motif (GGG-X6-8-QEE)10 and Tyr791 that contributes strongly to PARG catalytic efficiency and inhibitor binding. The structure consists of a alpha-beta-alpha fold with a nine-stranded, mixed beta-sheet flanked by several layers of alpha-helices	Rattus norvegicus

Synonyms

Synonyms	Comment	Organism
PARG	-	Rattus norvegicus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Rattus norvegicus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Rattus norvegicus

General Information

General Information	Comment	Organism
additional information	structure analysis and comparisons, overview. The poorly structured A-domain does not contribute to PARG activity in vitro. The rPARG385 catalytic domain adopts a beanshaped structure with a deep central cleft containing the conserved PARG-signature motif (GGG-X6-8-QEE)10 and Tyr791 that contributes strongly to PARG catalytic efficiency and inhibitor binding. The active site cleft lies on one edge of the beta-sheet and an extended N-terminal segment containing the MTS wraps around the other edge of the beta-sheet, contributing to the PARG catalytic domain	Rattus norvegicus
physiological function	poly(ADP-ribose) glycohydrolase catalyzes the removal of poly(ADP-ribose) chains from posttranslationally modified proteins by hydrolysis of alpha(122-22) O-glycosidic linkages, functioning as an endo-glycosidase to release oligo(ADP-ribose) and as an exo-glycosidase to release ADP-ribose	Rattus norvegicus