Literature summary for 3.6.1.13 extracted from

Huang, N.; Sorci, L.; Zhang, X.; Brautigam, C.A.; Li, X.; Raffaelli, N.; Magni, G.; Grishin, N.V.; Osterman, A.L.; Zhang, H.
Bifunctional NMN adenylyltransferase/ADP-ribose pyrophosphatase: structure and function in bacterial NAD metabolism (2008), Structure, 16, 196-209.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of GST-tagged nezyme in Escherichia coli	Francisella tularensis
expression of the His-tagged enzyme in Escherichia coli strain Bl21	Synechocystis sp.

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme in complex with the product AMP and Mn2+ ions in its Nudix active site, sitting drop vapor diffusion method, 20°C, 0.001 ml of 15 mg/ml protein solution is mixed with an equal volume of reservoir solution containing 0.1 M Tris, pH 7.5, 200 mM MgCl2, and 19% PEG 3350, with or without 30 mM AMP, equilibration against the reservoir, X-ray diffraction structure determination and analysis at 2.3 A resolution, molecular replacement method	Francisella tularensis
purified recombinant His-tagged enzyme in complex with co-purified NAD and diphosphate complexed in the NadM-domain active site, and with ADPR substrate complexed in the Nudix-domain, hanging drop vapor diffusion method, 0.0015 ml of 15 mg/ml protein solution is mixed with an equal volume of reservoir solution containing 100 mM Tris, pH 7.5, and 1.5 M Li2SO4, 20°C, 3 days to 2 weeks, X-ray diffraction structure determination and analysis at 2.6 A resolution, selenomethionyl MAD phasing method	Synechocystis sp.

Crystallization (Comment)

Organism

purified recombinant enzyme in complex with the product AMP and Mn2+ ions in its Nudix active site, sitting drop vapor diffusion method, 20°C, 0.001 ml of 15 mg/ml protein solution is mixed with an equal volume of reservoir solution containing 0.1 M Tris, pH 7.5, 200 mM MgCl2, and 19% PEG 3350, with or without 30 mM AMP, equilibration against the reservoir, X-ray diffraction structure determination and analysis at 2.3 A resolution, molecular replacement method

Francisella tularensis

purified recombinant His-tagged enzyme in complex with co-purified NAD and diphosphate complexed in the NadM-domain active site, and with ADPR substrate complexed in the Nudix-domain, hanging drop vapor diffusion method, 0.0015 ml of 15 mg/ml protein solution is mixed with an equal volume of reservoir solution containing 100 mM Tris, pH 7.5, and 1.5 M Li2SO4, 20°C, 3 days to 2 weeks, X-ray diffraction structure determination and analysis at 2.6 A resolution, selenomethionyl MAD phasing method

Synechocystis sp.

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Francisella tularensis
Mg2+	-	Synechocystis sp.

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ADP-ribose + H2O	Francisella tularensis	-	AMP + D-ribose 5-phosphate	-	?
ADP-ribose + H2O	Synechocystis sp.	-	AMP + D-ribose 5-phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Francisella tularensis	Q5NHR1	-	-
Synechocystis sp.	Q55928	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant GST-tagged enzyme from Escherichia coli by glutathione affinity chromatography, the tag is cleaved off by TEV protease, and the detagged protein is further purified by anion exchange chromatography	Francisella tularensis
recombinant His-tagged enzyme from Escherichia coli strain Bl21 by nickel affinity and anion exchange chromatography, and gel filtration	Synechocystis sp.

Reaction

Reaction	Comment	Organism	Reaction ID
ADP-D-ribose + H2O = AMP + D-ribose 5-phosphate	substrate binding mode in the active site of the ADPRase domain and catalytic mechanism	Francisella tularensis	a
ADP-D-ribose + H2O = AMP + D-ribose 5-phosphate	substrate binding mode in the active site of the ADPRase domain and catalytic mechanism	Synechocystis sp.	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ADP-ribose + H2O	-	Francisella tularensis	AMP + D-ribose 5-phosphate	-	?
ADP-ribose + H2O	-	Synechocystis sp.	AMP + D-ribose 5-phosphate	-	?
ADP-ribose + H2O	substrate binding structure, overview	Francisella tularensis	AMP + D-ribose 5-phosphate	-	?
ADP-ribose + H2O	substrate binding structure, overview	Synechocystis sp.	AMP + D-ribose 5-phosphate	-	?
additional information	the enzyme is a bifunctional NMN adenylyltransferase/ADP-ribose diphosphatase, NMN specificity of the enzyme, overview	Francisella tularensis	?	-	?
additional information	the enzyme is a bifunctional NMN adenylyltransferase/ADP-ribose diphosphatase, NMN specificity of the enzyme, overview	Synechocystis sp.	?	-	?

Subunits

Subunits	Comment	Organism
More	ADPRase Nudix domain of NadM-Nudix and bacterial monofunctional ADPRase are connected by a long alpha helix with ADPRase active site facing away from the NMNATase domain, while the NMNATase active site opens partially toward the ADPRase domain, structure of the ADPRase domain, overview	Francisella tularensis
More	ADPRase Nudix domain of NadM-Nudix and bacterial monofunctional ADPRase are connected by a long alpha helix with ADPRase active site facing away from the NMNATase domain, while the NMNATase active site opens partially toward the ADPRase domain, structure of the ADPRase domain, overview	Synechocystis sp.

Synonyms

Synonyms	Comment	Organism
NadM-Nudix	-	Francisella tularensis
NadM-Nudix	-	Synechocystis sp.
NMN adenylyltransferase/ADP-ribose pyrophosphatase	-	Francisella tularensis
NMN adenylyltransferase/ADP-ribose pyrophosphatase	-	Synechocystis sp.

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Francisella tularensis
37	-	assay at	Synechocystis sp.

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.2	-	assay at	Francisella tularensis
8.2	-	assay at	Synechocystis sp.