Refolding Record:
| Protein | |
|---|---|
| Protein Name | Indole 3-glycerol phosphate synthase |
| Abbreviated Name | IGPS |
| SCOP Family | Tryptophan biosynthesis enzymes |
| Structure Notes | |
| Organism | Escherichia coli |
| UniProt Accession | Q8X7B7 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha/Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | n |
| Domain | n/a |
| Chimera | n/a |
| Variants | IGPS mutant (49-252) |
| Chain Length | 207 |
| Molecular Weight | 23015.4 |
| Pi | 5.23 |
| Molecular Weight | 23015.4 |
| Disulphides | 0 |
| Full Sequence |
FILECKKASPSKGVICDDFDPARIAAIYKHYASAISVLTDEKYFQGSFDFLPIVSQIAPQPILCKDFIIDPYQIYLARYYQADACLLMLSVLDDEQYRQLAAVAHSLKMGVLTEVSNEEELERAIALGAKVVGINNRDLRDLSIDLNRTRELAPKLGHNVTVISESGINTYAQVRELSHFADGFLIGSALMAHDDLHAAVRRVLLGE
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Altamirano, M. M., Golbik, R., Zahn, R., Buckle, A. M., Fersht, A. R. (1997) Proc. Natl. Acad. Sci. USA, 94, 3576-3578 |
| Project Aim | Folding |
| Fusion | None |
| Protein Expression and Production | Protein expressed and purified in native conformation prior to denaturation and refolding. |
| Expression Host | None |
| Expression Strain | None |
| Expression Temp | 0.0 |
| Expression Time | n/a |
| Expression Vector | n/a |
| Expression Protocol | n/a |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Column refolding: Nickel-chelating chromatography |
| Wash Buffer | n/a |
| Solubilization Buffer | 8 M urea, 0.1 M potassium phosphate, pH 7.8, 5mM 2-mercaptoethanol |
| Refolding Buffer | 0.1 M potassium phosphate, pH 7.8, 5mM 2-mercaptoethanol |
| Pre-Refolding Purification | None |
| Tag Cleaved | no |
| Refolding pH | 7.8 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | A column of ht-GroEl 191-345 immobilized on Ni-NTa agarose and equilibrated with refolding buffer was loaded with 2 nmol of IGPS(49-242) in 8M urea. Refolded IGPS was then eluted as for conventional column. |
| Refolding Assay | Far-UV Circular Dichroism |
| Refolding Chaperones | GroEL minichaperone |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |