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Literature summary extracted from

  • Wojcik, M.; Seidle, H.F.; Bieganowski, P.; Brenner, C.
    Glutamine-dependent NAD+ synthetase. How a two-domain, three-substrate enzyme avoids waste (2006), J. Biol. Chem., 281, 33395-33402.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
6.3.5.1 nicotinic acid adenine dinucleotide stimulates activity 50fold Saccharomyces cerevisiae

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.1.5 expressed in Escherichia coli Saccharomyces cerevisiae
6.3.5.1 expressed in Escherichia coli Saccharomyces cerevisiae

Protein Variants

EC Number Protein Variants Comment Organism
6.3.1.5 D593A poor synthetase activity Saccharomyces cerevisiae
6.3.1.5 D593A/F622A synthetase dead Saccharomyces cerevisiae
6.3.1.5 E177A poor synthetase activity Saccharomyces cerevisiae
6.3.1.5 F622A poor synthetase activity Saccharomyces cerevisiae
6.3.1.5 I111A poor synthetase activity Saccharomyces cerevisiae
6.3.1.5 L529A/L604A poor synthetase activity Saccharomyces cerevisiae
6.3.1.5 L604A poor synthetase activity Saccharomyces cerevisiae
6.3.1.5 L604N poor synthetase activity Saccharomyces cerevisiae
6.3.1.5 M621A synthetase mutant that depresses all activities Saccharomyces cerevisiae
6.3.1.5 R112L poor synthetase activity that inhibit substrate synergism Saccharomyces cerevisiae
6.3.1.5 R112S poor synthetase activity that inhibit substrate synergism Saccharomyces cerevisiae
6.3.1.5 Y532A/Y601A synthetase dead Saccharomyces cerevisiae
6.3.1.5 Y601A poor synthetase activity Saccharomyces cerevisiae
6.3.1.5 Y601A/M621A synthetase dead Saccharomyces cerevisiae
6.3.5.1 D593A poor synthetase activity Saccharomyces cerevisiae
6.3.5.1 D593A/F622A synthetase dead Saccharomyces cerevisiae
6.3.5.1 E177A poor synthetase activity Saccharomyces cerevisiae
6.3.5.1 F622A poor synthetase activity Saccharomyces cerevisiae
6.3.5.1 I111A poor synthetase activity Saccharomyces cerevisiae
6.3.5.1 L529A/L604A poor synthetase activity Saccharomyces cerevisiae
6.3.5.1 L604A poor synthetase activity Saccharomyces cerevisiae
6.3.5.1 L604N poor synthetase activity Saccharomyces cerevisiae
6.3.5.1 M621A synthetase mutant that depresses all activities Saccharomyces cerevisiae
6.3.5.1 R112L poor synthetase activity that inhibit substrate synergism Saccharomyces cerevisiae
6.3.5.1 R112S poor synthetase activity that inhibit substrate synergism Saccharomyces cerevisiae
6.3.5.1 Y532A/Y601A synthetase dead Saccharomyces cerevisiae
6.3.5.1 Y601A poor synthetase activity Saccharomyces cerevisiae
6.3.5.1 Y601A/M621A synthetase dead Saccharomyces cerevisiae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.5.1 1.4
-
 L-glutamine mutant enzyme Y601A, at 37°C Saccharomyces cerevisiae
6.3.5.1 3.5
-
 L-glutamine mutant enzyme Y532A/Y601A, at 37°C Saccharomyces cerevisiae
6.3.5.1 5.3
-
 L-glutamine wild type enzyme, at 37°C Saccharomyces cerevisiae

Organism

EC Number Organism UniProt Comment Textmining
6.3.1.5 Saccharomyces cerevisiae
-
-
-
6.3.5.1 Saccharomyces cerevisiae
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.5.1
-
 Saccharomyces cerevisiae

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.3.1.5 0.0003
-
 mutant enzyme R112L, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.0011
-
 mutant enzyme D593A/F622A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.0027
-
 mutant enzyme L604A/L519A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.0033
-
 mutant enzyme Y532A/M621A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.0043
-
 mutant enzyme L604N, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.008
-
 mutant enzyme F622A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.0143
-
 mutant enzyme I111A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.015
-
 mutant enzyme R112S, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.0485
-
 mutant enzyme Y601A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.152
-
 mutant enzyme D593A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.183
-
 mutant enzyme L604A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.25
-
 mutant enzyme M621A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.631
-
 mutant enzyme E177A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.1.5 0.741
-
 wild type enzyme, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.0002
-
 mutant enzyme R112L, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.0011
-
 mutant enzyme D593A/F622A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.0017
-
 mutant enzyme Y532A/M621A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.0017
-
 mutant enzyme Y532A/Y601A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.0087
-
 mutant enzyme L604N, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.015
-
 mutant enzyme L604A/L519A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.024
-
 mutant enzyme R112S, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.0533
-
 mutant enzyme I111A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.0931
-
 mutant enzyme Y601A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.112
-
 mutant enzyme E177A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.288
-
 mutant enzyme F622A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.293
-
 mutant enzyme L604A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.435
-
 mutant enzyme D593A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 0.644
-
 mutant enzyme M621A, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae
6.3.5.1 2.33
-
 wild type enzyme, at 37°C, with 2 mM ATP, 5 mM MgCl2, 50 mM Tris-HCl, pH 8.0, 56 mM KCl, and 0.2 mg/ml bovine serum albumin Saccharomyces cerevisiae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.1.5 ATP + deamido-NAD+ + NH3
-
 Saccharomyces cerevisiae AMP + diphosphate + NAD+
-
 ?
6.3.5.1 ATP + deamido-NAD+ + L-glutamine + H2O
-
 Saccharomyces cerevisiae AMP + diphosphate + NAD+ + L-glutamate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.3.1.5 NH3-dependent NAD+ synthetase
-
 Saccharomyces cerevisiae
6.3.1.5 Qns1 couples a glutamine amidotransferase domain to a second active site that requires ammonia as a reactant Saccharomyces cerevisiae
6.3.5.1 glutamine-dependent NAD+ synthetase
-
 Saccharomyces cerevisiae
6.3.5.1 Qns1 couples a glutamine amidotransferase domain to a second active site that requires ammonia as a reactant Saccharomyces cerevisiae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.3.5.1 0.12
-
 L-glutamine mutant enzyme Y532A/Y601A, at 37°C Saccharomyces cerevisiae
6.3.5.1 0.16
-
 L-glutamine mutant enzyme Y601A, at 37°C Saccharomyces cerevisiae
6.3.5.1 6.4
-
 L-glutamine wild type enzyme, at 37°C Saccharomyces cerevisiae

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.1.5 ATP
-
 Saccharomyces cerevisiae
6.3.5.1 ATP
-
 Saccharomyces cerevisiae