Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Pseudomonas fluorescens |
Crystallization (Comment) | Organism |
---|---|
hanging-drop vapor-diffusion method. Crystals of unliganded PhzF belong to space group P3(2)21 with a a = b = 56 A, and c = 156 A and hold one monomer per asymmetric unit. Complexes of PhzF (with 3-hydroxyanthranilic acid and trans-2,3-dihydro-3-hydroxyanthranilic acid) crystallize in space group P2(1)2(1)2 with cell parameters of a = 93 A, b = 100 A, and c = 57 A and contain one dimer in the asymmetric unit | Pseudomonas fluorescens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
42000 | - |
gel filtration | Pseudomonas fluorescens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5S,6S)-6-amino-5-hydroxycyclohexane-1,3-diene-1-carboxyate | Pseudomonas fluorescens | i.e. trans-2,3-dihydro-3-hydroxyanthranilic acid. The enzyme is involved in phenazine synthesis | (1R,6S)-6-amino-5-oxocyclohex-2-ene-1-carboxylate | i.e. 2,3-dihydro-3-hydroxy-anthranilate | ? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas fluorescens | Q51792 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas fluorescens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5S,6S)-6-amino-5-hydroxycyclohexane-1,3-diene-1-carboxyate | i.e. trans-2,3-dihydro-3-hydroxyanthranilic acid. The enzyme is involved in phenazine synthesis | Pseudomonas fluorescens | (1R,6S)-6-amino-5-oxocyclohex-2-ene-1-carboxylate | i.e. 2,3-dihydro-3-hydroxy-anthranilate | ? | |
(5S,6S)-6-amino-5-hydroxycyclohexane-1,3-diene-1-carboxyate | charactierzation of the product by NMR spectrometry. The structure of PhzF in complex with its substrate, trans-2,3-dihydro-3-hydroxyanthranilic acid, suggests that it is an isomerase using the conserved glutamate45 to abstract a proton from C3 of the substrate. The proposed mechanism involves proton abstraction from C3 of trans-2,3-dihydro-3-hydroxyanthranilate by Glu45, followed by rearrangement of the double bonds and reprotonation at C1. The function of E45 as a proton shuttle is supported by results from NMR spectroscopy | Pseudomonas fluorescens | (1R,6S)-6-amino-5-oxocyclohex-2-ene-1-carboxylate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | deduced from crystal structure. The crystal structure clarifies that the enzyme is a dimer in an up/up configuration with the two active sites facing each other. The dimer in the open form, generates by crystal symmetry (sulfate complex), interacts mainly through the alpha1-helices and the beta16-strands in the N-terminal domains. The C-terminal domains are not in direct contact but are relatively close | Pseudomonas fluorescens |
Synonyms | Comment | Organism |
---|---|---|
phzF | - |
Pseudomonas fluorescens |