Refolding Record:
| Protein | |
|---|---|
| Protein Name | Interferon gamma |
| Abbreviated Name | IFN gamma |
| SCOP Family | Interferons/interleukin-10 (IL-10) |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P01579 |
| 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 | 146 |
| Molecular Weight | 17145.6 |
| Pi | 9.54 |
| Molecular Weight | 17145.6 |
| Disulphides | 0 |
| Full Sequence |
CYCQDPYVKEAENLKKYFNAGHSDVADNGTLFLGILKNWKEESDRKIMQSQIVSFYFKLFKNFKDDQSIQKSVETIKEDMNVKFFNSNKKKRDDFEKLTNYSVTDLNVQRKAIHELIQVMAELSPAAKTGKRKRSQMLFRGRRASQ
|
| Notes | Full length without signal sequence (1-20). |
| Expression | |
|---|---|
| Report | Arakawa, T., Alton, N. K., Hsu, Y. R (1985) J Biol Chem, 260, 14435-14439 |
| 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 | Dialysis |
| Wash Buffer | n/a |
| Solubilization Buffer | 7 M Urea |
| Refolding Buffer | 0.1 M Nh4OAc, I M urea |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.5 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | 7 M Solid urea was added to purified IFNG (1 mg/ml in 0.1 M NH4OAc) to unfold protein. A two day dialysis was preformed after a two hour incubation period which followed a complete dissolution of the added urea. The solution was applied to a Sephadex G-75 column in the same buffer. The two peaks produced underwent rechromatographying separately. |
| Refolding Assay | Far-UV Circular Dichroism |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | IFN-Gamma exists in reversible equilibrium between monomer and oligomer in refolding buffer. |