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Literature summary extracted from

  • Morandi, P.; Valzasina, B.; Colombo, C.; Curti, B.; Vanoni, M.A.
    Glutamate Synthase: Identification of the NADPH-Binding Site by Site-Directed Mutagenesis (2000), Biochemistry, 39, 727-735.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.1.13 production of the G298A-beta subunit in Escherichia coli Azospirillum brasilense

Protein Variants

EC Number Protein Variants Comment Organism
1.4.1.13 G298A mutant with an approximately 10fold decrease of the affinity of the enzyme for pyridine nucleotides with little or no effect on the rate of the enzyme reduction by NADPH, maintains the ability to bind NADPH and FAD, is catalytically active in the NADPH:iodonitrotetrazolium oxidoreductase reaction and has a monomeric state, mutation leads to production of insoluble protein under conditions that yield large amounts of soluble wild-type enzyme and to production of smaller amounts of enzyme Azospirillum brasilense

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.4.1.13 2'-phosphoadenosine-5'-diphospho-5'-beta-D-ribose inhibitor of the NADPH:iodonitrotetrazolium oxidoreductase reaction of the G298A-beta subunit, competitive with respect to NADPH Azospirillum brasilense
1.4.1.13 3-Aminopyridine adenine dinucleotide phosphate inhibitor of the NADPH:iodonitrotetrazolium oxidoreductase reaction of the G298A-beta subunit, competitive with respect to NADPH Azospirillum brasilense
1.4.1.13 NADP+ inhibitor of the NADPH:iodonitrotetrazolium oxidoreductase reaction of the G298A-beta subunit, competitive with respect to NADPH Azospirillum brasilense

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.1.13 0.084
-
 NADPH of NADPH: iodonitrotetrazolium oxidoreductase activity of the G298A-beta subunit Azospirillum brasilense

Organism

EC Number Organism UniProt Comment Textmining
1.4.1.13 Azospirillum brasilense
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.1.13 of the G298A-beta subunit, using column chromatography on Reactive Red or Amicon Red resins and gel filtration on Ultrogel AcA 34 Azospirillum brasilense

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.1.13 L-glutamine + 2-oxoglutarate + NADPH + H+
-
 Azospirillum brasilense L-glutamate + NADP+
-
 ?

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.1.13 flavin 0.86 mol FAD per mol of mutant beta subunit, 0.83 mol FAD per mol of the wild type species Azospirillum brasilense
1.4.1.13 NADPH the C-terminal potential ADP-binding fold of the beta subunit is the NADPH-binding site of the enzyme Azospirillum brasilense

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.4.1.13 0.0112
-
 3-Aminopyridine adenine dinucleotide phosphate inhibitor of the NADPH: iodonitrotetrazolium oxidoreductase reaction of wild type-beta subunit Azospirillum brasilense
1.4.1.13 0.0227
-
 2'-phosphoadenosine-5'-diphosphoribose inhibitor of the NADPH: iodonitrotetrazolium oxidoreductase reaction of wild type-beta subunit Azospirillum brasilense
1.4.1.13 0.037
-
 NADP+ inhibitor of the NADPH: iodonitrotetrazolium oxidoreductase reaction of G298A-beta subunit Azospirillum brasilense
1.4.1.13 0.128
-
 3-Aminopyridine adenine dinucleotide phosphate inhibitor of the NADPH: iodonitrotetrazolium oxidoreductase reaction of G298A-beta subunit Azospirillum brasilense
1.4.1.13 0.233
-
 2'-phosphoadenosine-5'-diphosphoribose inhibitor of the NADPH: iodonitrotetrazolium oxidoreductase reaction of G298A-beta subunit Azospirillum brasilense