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4.2.1.20: tryptophan synthase

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

Word Map on EC 4.2.1.20

Reaction

1-C-(indol-3-yl)glycerol 3-phosphate
=
indole
 +
D-glyceraldehyde 3-phosphate

Synonyms

alpha2beta2 tryptophan synthase, alphaTS, AtTSB1, beta subunit of tryptophan synthase, indoleglycerol phosphate aldolase, It-TSA, L-serine hydro-lyase (adding indoleglycerol-phosphate), L-tryptophan synthetase, PtTSA, Rv1612, synthase, tryptophan, TrB, Trp synthase, Trp synthase beta, TrpA, trpB, TrpB1, TrpB2, TrpB2a, TrpB2i, TrpB2o, TRPS, tryptophan desmolase, tryptophan synthase, tryptophan synthase alpha subunit, tryptophan synthase alpha-subunit, tryptophan synthase alpha2beta2 complex, tryptophan synthase beta, tryptophan synthase beta 1, tryptophan synthase beta subunit, tryptophan synthetase, TS, TSA, TSase, TSB, TSB1, TSbeta

ECTree

     4 Lyases
         4.2 Carbon-oxygen lyases
             4.2.1 Hydro-lyases
                4.2.1.20 tryptophan synthase

Crystallization

Crystallization on EC 4.2.1.20 - tryptophan synthase

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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
10 mg/ml purified recombinant alpha-subunit, hanging drop vapour diffusion method, 298K, equal volume, 0.001 ml, of protein solution and reservoir solution are mixed and placed over 0.5 ml reservoir solution, precipitant solution: 0.5 M ammonium sulfate, 0.1 M trisodium citrate dihydrate, 1.0 M lithium sulfate monohydrate, pH 5.6, first crystals after 7-10 days, maximal size within 2 weeks, X-ray diffraction structure determination and analysis at 2.8 A resolution
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crystal structures of apo-beta2 and holo-beta2 from Escherichia coli is determined at 3.0 and 2.9 A resolutions. The apo-type and holo-type molecule retain a dimeric form in solution. The subunit structures of both the apo-beta2 and the holo-beta2 forms consist of two domains, (N domain, C domain). The pyridoxal 5’-phosphate-bound holo-form has multiple interactions between the two domains and a long loop (residues 260-310), which are missing in the apo-form
crystal structures of wild-type and mutant P38L/Y173F alpha-subunit, 2.8 A and 1.8 A resolution, hanging-drop vapour diffusion method
hanging drop vapour diffusion method. Crystal structure of the tryptophan synthase alpha-subunit determined at 2.3 A resolution. Structure of tryptophan synthase alpha-subunit from Escherichia coli is compared to structure of alpha2beta2 complex from Salmonella typhimurium
wild-type and P28L/Y173F double mutant alpha-subunits are crystallized at 25°C by the hanging-drop vapor-diffusion method. X-ray diffraction data are collected to 2.5 A resolution from the wild-type crystals and to 1.8 A from the crystals of the double mutant. The wild-type crystals belonged to the monoclinic space group C2 (a = 155.64 A, b = 44.54 A, c = 71.53 A and beta = 96.39°) and the P28L/Y173F crystals to the monoclinic space group P 2(1) (a = 71.09 A, b = 52.70 A, c = 71.52 A, and beta = 91.49°). The asymmetric unit of both structures contains two molecules of tryptophan synthase alpha-subunit
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crystal structure of tryptophan synthase beta2 subunit determined at 2.2 A resolution, hanging drop vapour diffusion method. Crystals belong to the orthorhombic space group P2(1)2(1)2(1) with unit cell dimensions of a = 84.8 A, b = 110.5 A, c = 160.0 A
hanging drop vapour diffusion method, crystal structure of the alpha2beta2 complex, crystals belong to the orthorhombic space group of P2(1)2(1)2(1) with unit cell dimensions of a = 89.1 A, b = 220.3 A, c = 292.6 A
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purified recombinant alpha-subunit, hanging drop vapour diffusion method, 10°C, reservoir solution: 0.1 M MES-NaOH, pH 6.5, 12% PEG 20000, X-ray diffraction structure determination and analysis at 2.0 A resolution
wild-type in complex with L-threonine, to 1.54 A. L-threonine binds non-covalently, no formation of an electrophilic amino-acrylate intermediate. With mutant I16V/E17G/I68V/F95L/F274S/T292S/T321A/V384A, the species is formed
crystal structures of the alpha-subunit of tryptophan synthase alone and in the alpha2beta2 complex
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crystallization of wild-type and mutant S178P in presence or absence of alpha-subunit ligands, structure comparison
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crystallization of wild-type enzyme alone and complexed with L-serine and 3-indolylpropanol 3'-phosphate, and of mutant T183V complexed with L-serine, 1-(indol-3-yl)glycerol 3-phosphate, and 3-indolylpropanol 3'-phosphate, hanging drop method in the dark at room temperature, equal volumes of protein solution, containing 10 mg/ml protein, 50 mM Na-bicine, pH 7.8, 10 mM Na-EDTA, 1 mM dithioerythritol, 0.02 mM pyridoxal 5'-phosphate, and of reservoir solution, containing 50 mM Na-bicine, pH 7.8, 5 mM dithioerythritol, 5 mM Na-EDTA, 0.1 mM pyridoxal 5'-phosphate, 2 mM spermine, 8-12% w/v PEG 8000, X-ray diffraction structure determination and analysis at different resolutions 1.45-2.3 A, comparison of crystal structures of the different complexes
-
hanging drop method, 10 mg/ml purified recombinant mutant S178P enzyme in 50 mM bicine, pH 7.8, with equal volume of reservoir solution: 50 mM bicine, 1 mM EDTA, 12% w/v PEG 8000, 1.4 mM spermine, pH 7.8, 21°C, structure determination and functional investigation by polarized absorption microspectrophotometry
-
hanging drop vapour diffusion method
-
in complex with inhibitor N-(4'-trifluoro-methoxybenzenesulfonyl)-2-amino-ethyl phosphate
molecular dynamics simulations. The unprotonated pyridine nitrogen can form an H-bond with Ser377, which stabilizes the PLP ring structure. When the pyridoxyl phenolic oxygen is unprotonated, the polar side chain of Gln114 moves toward the negatively charged oxygen, which affects the movements of the water molecules around the active site and induces the switch to the open conformation of the beta subunit. The carboxylate oxygens typically form stable H-bonds with Thr110, Gly111, and His115
purified enzyme complexed with allosteric effectors indole-3-acetylglycine and indole-3-acetyl-L-aspartic acid, hanging drop method, in the dark at room temperature, equal volumes of protein and reservoir solution, the latter containing 9-12% PEG 8000, 1.5 mM spermine, 1 mM EDTA, 50 mM bicine, pH 7.8, X-ray diffraction structure determination and analysis at 2.5 A resolution, modeling
-
purified mutant A169L/C170W complexed with the alpha-active site substrate analogue 5-fluoro-indole-propanol-phosphate, X-ray diffraction structure determination and analysis at 2.25 A resolution
-
purified recombinant enzyme complex, structure determination and analysis
-
purified recombinant enzyme in complex with potential alpha-reaction phosphonate inhibitors, 5-10 mg/ml protein mixed with 1 mM inhibitor, in 50 mM bicine, 1 mM Na-EDTA, 0.8-1.5 mM spermine, 12% PEG 4000, to pH 7.8 with NaOH, X-ray diffraction structure determination and analysis at 2.3 A resolution or higher
-
purified recombinant wild-type enzyme alone or in complex with alpha-subunit substrate analogue indole propanol phosphate, 10 mg/ml in 50 mM bicine, pH 7.8, 10 mM Na-EDTA, 0.02 mM pyridoxal 5'-phosphate, 1 mM dithioerythritol, in the dark at room temperature, hanging drop method, equal volumes of protein and reservoir solution, the latter containing 50 mM bicine, pH 7.8, 5 mM dithioerytritol, 5 mM Na-EDTA, 0.1 mM pyridoxal 5'-phosphate, 2 mM spermine, 2 mM NaN3, and 8-12% w/v PEG 8000, with or without 7 mM indole propanol phosphate, X-ray diffraction structure determination and analysis at 1.4 A resolution, structure modeling
-
single-molecule stochastic model of the enzyme, with the transition state constants deduced from the experimental data available. The model yields direct theoretical evidence for intramolecular synchronization phenomena
soaking native crystals for 10 min in a solution containing 90 mM Bis-Tris-propane (pH 7.8), 150 mM NaCl, 15% (w/v) PEG 8000, and 20% glycerol
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structure and function of alpha subunit, beta subunit, alpha2beta2 complex
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structures of tryptophan synthase crystallized with varying numbers of amphipathic N-(4'-trifluoromethoxybenzoyl)-2-aminoethyl phosphate. One of the ligands threads into the tunnel from the beta-site and reveals a distinct hydrophobic region. The N-(4'-trifluoromethoxybenzoyl)-2-aminoethyl phosphate phosphoryl group fits into a polar pocket of the beta-subunit active site. One portion of the tunnel binds clusters of water molecules, whereas waters are not observed in the nonpolar N-(4'-trifluoromethoxybenzoyl)-2-aminoethyl phosphate binding region of the tunnel. Hydrophobic molecules can freely diffuse between the alpha- and beta-sites via the tunnel, while water does not. Exclusion of water may serve to inhibit reaction of water with the alpha-aminoacrylate intermediate to form ammonium ion and pyruvate
vapor diffusion method, using 1 mM dithiothreitol, 0.04% (w/v) NaN3, and 12% (w/v) polyethylene glycol 8000
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sitting drop vapor diffusion method, using 0.1M HEPES-Na, pH 7.5, 2% (v/v) polyethylene glycol 400 and 2 M ammonium sulfate, at 4°C
-
crystal structure of alpha-subunit of tryptophan synthase, oil batch method, discussion of the thermostabilization mechanism of the tryptophan synthase alpha-subunit on the basis of crystal structures and DSC data of the alpha-subunit orthologs from mesophilic, extreme thermophilic, and hyperthermophilic organisms