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4.1.99.1: tryptophanase

This is an abbreviated version!
For detailed information about tryptophanase, go to the full flat file.

Word Map on EC 4.1.99.1

Reaction

L-tryptophan
+
H2O
=
indole
 +
pyruvate
 +
NH3

Synonyms

L-tryptophan indole-lyase, L-tryptophanase, TIL, tna2, TnaA, tnaA2, TNase, Tpase, Trpase, tryptophan indole lyase, tryptophan indole-lyase, tryptophan-indole lyase, tryptophanase, tryptophanase 2, VcTrpase

ECTree

     4 Lyases
         4.1 Carbon-carbon lyases
             4.1.99 Other carbon-carbon lyases
                4.1.99.1 tryptophanase

Crystallization

Crystallization on EC 4.1.99.1 - tryptophanase

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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
apo-form of enzyme without bound pyridoxal 5’-phosphate but with two bound sulfate ions, hanging drop vapor diffusion method
apo-form of the enzyme, hanging drop vapor diffusion method
-
crystallized in the apo form by the hanging-drop vapour-diffusion method using polyethylene glycol 400 as a precipitant and magnesium chloride as an additive. The crystals belong to the orthorhombic space group F222, with unit-cell parameters a = 118.4 A, b = 120.1 A, c = 171.2 A. Contains a monomer in the asymmetric unit with a solvent content of 55%. Tryptophanase mutants W330F and Y74F are crystallized under the same conditions and the crystals diffracted to a resolution limit of 1.9 A
-
mutant enzymes Y74F and C298S, hanging drop vapor diffusion method, using 30% (w/v) PEG 400, 100 mM HEPES pH 7.5, 200 mM MgCl2, 5 mM 2-mercaptoethanol
purified recombinant apo-enzyme in an open conformation, hanging drop vapour diffusion method with 25-30% PEG 4000, 30 mM ammonium sulfate, 20 mM 2-mercaptoethanol, 100 mM Tris-HCl, pH 9.0, X-ray diffraction structur determination and analysis at 2.8 A resolution
purified recombinant tryptophanase in holo- and semi-holoforms, hanging drop vapour diffusion method, mixing of 15-25 mg/ml protein in 50 mM sodium phosphate, pH 6.0, 5 mM 2-mercaptoethanol, and 0.5-2 mM pyridoxal 5'-phosphate in a ratio of 3:1 with precipitant solution containing 10-15% w/v ammonium sulfate, 25 mM potassium acetate, pH 5.4, and 3-10 mM2-mercaptoethanol, 2-7 days, X-ray diffraction structure determination and analysis at 2.9-3.2 A resolution, both crystal forms belong to the same space group P43212 but have slightly different unit-cell parameters, molecular replacement using the structure of apo Trpase, PDB ID 2oqx, as the search model
strong crystal contacts occur on the flat surface of the protein and that the size of crystal contact surface seems to correlate with the diffraction quality of the crystal. The tryptophanase structure, solved in its apo form, does not have covalent PLP bound in the active site, but two sulfate ions. The sulfate ions occupy the phosphoryl-binding site of PLP and the binding site of the alpha-carboxyl of the natural substrate tryptophan. One of the sulfate ions makes extensive interactions with both the transferase and PLP-binding domains of the protein and appears to be responsible for holding the enzyme in its closed conformation
analysis of the enzyme tetramer with one pyridoxal 5'-phosphate bound to each monomer, crystal structure at 2.1 A resolution, PDB ID 1AX4
in complex with oxindolyl-L-alanine and with L-tryptophan and L-serine
-
purified recombinant enzyme in complex with inhibitor oxindolyl-L-alanine (OIA), hanging-drop vapor diffusion method by mixing 15 mg/ml protein with an equal volume of reservoir solution consisting of 35 mM potassium phosphate, 0.05 M HEPES, pH 7.0, 0.3 M KCl, and 11% PEG 4000, at 20°C, the crystals are transferred to reservoir solution supplemented with 20% 1:1:1 ethylene glycol:DMSO:glycerol as cryosolvent, with or without 10 mM OIA, X-ray diffraction structure determination and analysis at 2.0-2.1 A resolution