Refolding Record:
| Protein | |
|---|---|
| Protein Name | Interleukin-2 |
| Abbreviated Name | IL-2 |
| SCOP Family | Short Chain Cytokines |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P60568 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 155 |
| Molecular Weight | 17627.7 |
| Pi | 7.6656 |
| Molecular Weight | 17627.7 |
| Disulphides | 1 |
| Full Sequence |
MYRMQLLSCIALSLALVTNSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRML
TFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSE
TTFMCEYADETATIVEFLNRWITFCQSIISTLT
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Tsuji T, Nakagawa R, Sugimoto N, Fukuhara K. (1987) Biochemistry, 26, 3129-3134 |
| Project Aim | Undefined |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21 |
| Expression Temp | 37.0 |
| Expression Time | unknown |
| Expression Vector | unknown |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | Lysozyme |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution/Column Refolding Combination |
| Wash Buffer | unknown |
| Solubilization Buffer | 100mM Tris-HCl, 6M guanidinium chloride, pH 8.0 |
| Refolding Buffer | 100mM Tris-HCl, 10mM GSH, 1mM GSSG, 2M guanidinium chloride |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | 0.1mg/ml |
| Refolding Time | 16h |
| Redox Agent | GSH/GSSG |
| Redox Agent Concentration | n/a,n/a |
| Refolding Protocol | The denatured inclusion bodies were diluted into buffer such that the protein concentration was 0.1mg/ml and the guanidinium chloride concentration was 2M. The solution was treated with 10mM reduced glutathione and 1mM oxidised glutathione and was kept at room temperature for 16h. Subsequently the solution was desalted on a Sephadex G25 column equlibrated with 50mM sodium acetate pH 6.0. |
| Refolding Assay | Unspecified |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | |
| Notes | |