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Literature summary extracted from

  • Ghosh, J.; Anderson, P.J.; Chandrasekaran, S.; Caparon, M.G.
    Characterization of Streptococcus pyogenes beta-NAD+ glycohydrolase: re-evaluation of enzymatic properties associated with pathogenesis (2010), J. Biol. Chem., 285, 5683-5694.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.2.5 recombinant expression of His-tagged wild-type and E391Q mutant SPN in Escherichia coli as stable and soluble monomer in the periplasmic space Streptococcus pyogenes

Protein Variants

EC Number Protein Variants Comment Organism
3.2.2.5 E391Q a catalytically nearly inactive mutant, upon transformation, SPN is toxic to Saccharomyces cerevisiae, whereas a glycohydrolase-inactive SPN allows for viability Streptococcus pyogenes

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.2.5 ADP-ribose product inhibition Streptococcus pyogenes

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.2.5 additional information
-
 additional information Michaelis-Menten kinetics, and kinetic analysis Streptococcus pyogenes
3.2.2.5 0.188
-
 NAD+ pH 7.4, 37°C, recombinant wild-type enzyme Streptococcus pyogenes
3.2.2.5 0.294
-
 NAD+ pH 7.4, 37°C, recombinant mutant enzyme Streptococcus pyogenes

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.2.5 additional information Streptococcus pyogenes SPN is specific for beta-NAD+ glycohydrolase activity, and does not show ADP-ribosyl cyclase and ADP-ribosyltransferase activities, overview. SPN is unable to catalyze cyclic ADPR hydrolysis, and cannot catalyze methanolysis or transglycosidation ?
-
 ?
3.2.2.5 NAD+ + H2O Streptococcus pyogenes
-
 ADP-ribose + nicotinamide
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.2.5 Streptococcus pyogenes
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.2.5 recombinant His-tagged wild-type and E391Q mutant SPN from Escherichia coli Streptococcus pyogenes

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.2.5 NAD+ + H2O = ADP-D-ribose + nicotinamide + H+ SPN shows an ordered uni-bi mechanism, with ADP-ribose being released as a second product. The catalytic mechanism requires an essential glutamic acid, Glu344, to stabilize an oxycarbenium ion intermediate, which subsequently is attacked by a nucleophile, a water molecule Streptococcus pyogenes
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.2.2.5 additional information the Gram-positive pathogen Streptococcus pyogenes injects a beta-NAD+-glycohydrolase into the cytosol of an infected host cell using the cytolysin-mediated translocation pathway Streptococcus pyogenes
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.2.5 additional information SPN is specific for beta-NAD+ glycohydrolase activity, and does not show ADP-ribosyl cyclase and ADP-ribosyltransferase activities, overview. SPN is unable to catalyze cyclic ADPR hydrolysis, and cannot catalyze methanolysis or transglycosidation Streptococcus pyogenes ?
-
 ?
3.2.2.5 NAD+ + H2O
-
 Streptococcus pyogenes ADP-ribose + nicotinamide
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.2.5 beta-NAD+ glycohydrolase
-
 Streptococcus pyogenes
3.2.2.5 SPN
-
 Streptococcus pyogenes

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.2.5 37
-
 assay at Streptococcus pyogenes

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.2.5 2.2
-
 NAD+ pH 7.4, 37°C, recombinant mutant enzyme Streptococcus pyogenes
3.2.2.5 8390
-
 NAD+ pH 7.4, 37°C, recombinant wild-type enzyme Streptococcus pyogenes

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.2.5 7.4
-
 assay at Streptococcus pyogenes

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.2.5 3.6
-
 ADP-ribose pH 7.4, 37°C, recombinant wild-type enzyme Streptococcus pyogenes

General Information

EC Number General Information Comment Organism
3.2.2.5 additional information the Gram-positive pathogen Streptococcus pyogenes injects a beta-NAD+-glycohydrolase into the cytosol of an infected host cell using the cytolysin-mediated translocation pathway, where SPN accelerates the death of the host cell Streptococcus pyogenes