L-Ser, L-His, indoline, CsCl, NaCl, and NH4Cl were purchased from Sigma-Aldrich. Indoline was purified as previously described (17 (link)). N-(4-trifluoromethyoxylbenzenesulfonyl)-2-amino-1-ethylphosphate (F9) was synthesized as described (22 (link)). 5,5'-Dithio-bis(2-nitrobenzoic acid) (DTNB) was purchased from Calbiochem. All solutions were prepared at 25 ± 2° C and maintained at pH 7.8 in 50 mM triethanolamine (TEA) buffer and (except for H+) maintained MVC-free. F9 was prepared free of monovalent metal ions by exchange of Na+ with cyclohexylammonium chloride. Salmonella tryphimurium α2β2 tryptophan synthase was purified as previously described (28 (link)).
Protocol detail
Preparation and Purification of Tryptophan Synthase
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Buffer
Chloride
Cscl
Dtnb
Indoline
Ions
Metal
Nacl
Nitrobenzoic acid
Salmonella
Triethanolamine
Tryptophan synthase
Corresponding Organization :
Other organizations : University of California, Riverside, Max Planck Institute for Medical Research
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Variable analysis
independent variables
- L-Ser
- L-His
- Indoline
- NH4Cl
- N-(4-trifluoromethyoxylbenzenesulfonyl)-2-amino-1-ethylphosphate (F9)
- Not explicitly mentioned
- PH 7.8 in 50 mM triethanolamine (TEA) buffer
- Maintained MVC-free (except for H+)
- F9 prepared free of monovalent metal ions by exchange of Na+ with cyclohexylammonium chloride
- Salmonella tryphimurium α2β2 tryptophan synthase
- Not explicitly mentioned
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