Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Wang, M.; Yuan, Z.; Xie, R.; Ma, Y.; Liu, X.; Yu, X.
    Structure-function analyses reveal the mechanism of the ARH3-dependent hydrolysis of ADP-ribosylation (2018), J. Biol. Chem., 293, 14470-14480 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.143 gene ADPRHL2, sequence comparisons, recombinant expression of N-terminally His-tagged wild-type enzyme and GST-tagged mutant enzymes in Escherichia coli strain BL21(DE3) Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.143 purified ARH3-ADPR substrate complex, sitting drop vapour diffusion method, ARH3 and ADPR are mixed in the molar ratio of 1:3, mixing of equal volumes of 11 mg/ml protein complex in 10 mM Tris-HCl, pH 8.0, and 100 mM NaCl with reservoir solution consisting of 0.1 M MES, pH 6.0, and 20% w/v PEG MME 2000, addition of 0.2 M non-detergent sulfobetaine NDSB-201, 20°C, X-ray diffraction structure determination and analysis at 1.58 A resolution, molecular replacement using the structure of apo-ARH3 (PDB ID 2FOZ) as the search model, modeling Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.143 D314A site-directed mutagenesis, the mutation impairs ARH3-dependent DNA damage repair Homo sapiens
3.2.1.143 D77N site-directed mutagenesis, the mutation impairs ARH3-dependent DNA damage repair Homo sapiens
3.2.1.143 E41Q site-directed mutagenesis, the mutation impairs ARH3-dependent DNA damage repair Homo sapiens
3.2.1.143 H182A site-directed mutagenesis, the mutation impairs ARH3-dependent DNA damage repair Homo sapiens
3.2.1.143 additional information enzyme knockout by expression of ARH3 siRNA in U2OS cells Homo sapiens
3.2.1.143 S148A site-directed mutagenesis, the mutation impairs ARH3-dependent DNA damage repair Homo sapiens
3.2.1.143 T317A site-directed mutagenesis, the mutation impairs ARH3-dependent DNA damage repair Homo sapiens
3.2.1.143 Y149A site-directed mutagenesis, the mutation impairs ARH3-dependent DNA damage repair Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.143 Mg2+ required for catalysis, binuclear Mg2+ site, structure overview. The distal ribose of poly(ADP-ribose), enabling the hydroxyl groups of the distal ribose to coordinate the two Mg2+ ions. MgA is coordinated with Asp314, Asp316, Thr317, three hydroxyl groups of the distal ribose and water 269. Meanwhile, MgB is coordinated with the 2'-hydroxyl group of the distal ribose, Thr76, Asp77, Asp78, Asp316, and two water molecules (water 269 and water 85). The sequence alignment and structural comparison reveals that the Mg2+ coordinating acidic residues Asp77, Asp78, Asp314, and Asp316 are fully conserved in homologous ARH enzymes. Both Mg2+ ions are involved in the positioning of water 351 in the catalytic center through hydrogen-bonding interaction. the binding pocket of ARH3 is divided into two subunits by the binuclear Mg2+ sites Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.143 poly(ADP-ribose) + H2O Homo sapiens
-
 ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.143 Homo sapiens Q9NX46
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.143 recombinant His-tagged wild-type enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage through rhinovirus 3C (HRV 3C) protease, anion exchange chromatography, gel filtration, and ultrafiltration, recombinant GST-tagged mutant enzymes by glutathione affinity chromatography Homo sapiens

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.1.143 (ADP-ribose)n + H2O = (ADP-ribose)n-1 + ADP-ribose reaction mechanism and structure, overview Homo sapiens
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.143 poly(ADP-ribose) + H2O
-
 Homo sapiens ?
-
 ?
3.2.1.143 poly(ADP-ribose) + H2O enzyme-substrate binding complex structure analysis, overview Homo sapiens ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.143 More secondary structure comparisons, overview Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
3.2.1.143 ADPRHL2
-
 Homo sapiens
3.2.1.143 ARH3
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.143 22
-
 assay at room temperature Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.143 7.8
-
 assay at Homo sapiens

General Information

EC Number General Information Comment Organism
3.2.1.143 additional information analysis of the catalytic site structure of ARH3, overview Homo sapiens
3.2.1.143 physiological function enzyme ARH3 is a multifunctional enzyme that also hydrolyzes poly(ADP-ribose) (ADPR). Enzyme ARH3 plays a role in DNA damage repair. The recruitment of ARH3 to DNA lesions is mediated by ADPR recognition. The catalytic mechanism of protein ADP-ribose hydrolases can be classified into two different groups, namely metal-dependent and metal-independent catalysis. ARHs, such as ARH3, belong to metal-dependent catalysis, utilizing two Mg2+ ions and acidic residues to complete the catalytic reaction, which might be highly conserved. In contrast, the catalytic mechanism is not conserved in the macrodomain ADP-ribose hydrolases, For example, Glu756 and a water molecule act together to catalyze the reaction in PARG, whereas the key catalytic factor in MacroD2 is an activated water. The charge characteristic of the binding pocket in ARH3 is remarkably distinguished from that in PARG. The binding pocket of PARG, accommodating the ADPR dimer, is mostly composed of the basic region Homo sapiens