Refolding Record:
| Protein | |
|---|---|
| Protein Name | Interleukin-1 beta-converting enzyme |
| Abbreviated Name | ICE |
| SCOP Family | Caspase catalytic domain |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P29466 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha/Beta |
| Molecularity | Heterodimer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 410 |
| Molecular Weight | 45158.6 |
| Pi | 5.63287 |
| Molecular Weight | 45158.6 |
| Disulphides | 1 |
| Full Sequence |
MADKVLKEKRKLFIRSMGEGTINGLLDELLQTRVLNKEEMEKVKRENATVMDKTRALIDS
VIPKGAQACQICITYICEEDSYLAGTLGLSADQTSGNYLNMQDSQGVLSSFPAPQAVQDN
PAMPTSSGSEGNVKLCSLEEAQRIWKQKSAEIYPIMDKSSRTRLALIICNEEFDSIPRRT
GAEVDITGMTMLLQNLGYSVDVKKNLTASDMTTELEAFAHRPEHKTSDSTFLVFMSHGIR
EGICGKKHSEQVPDILQLNAIFNMLNTKNCPSLKDKPKVIIIQACRGDSPGVVWFKDSVG
VSGNLSLPTTEEFEDDAIKKAHIEKDFIAFCSSTPDNVSWRHPTMGSVFIGRLIEHMQEY
ACSCDVEEIFRKVRFSFEQPDGRAQMPTTERVTLTRCFYLFPGH
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Talanian et al (1996) J Biol Chem, 271, 21853-21858 |
| Project Aim | Crystallography |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | CAG597 |
| Expression Temp | 42.0 |
| Expression Time | 4h |
| Expression Vector | pL |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Microfluidizer |
| Lytic Agent | None |
| Pre-Refolding Purification | Reverse phase chromatography |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | see JBC 271:21853 for detailed protocol |
| Solubilization Buffer | see publication |
| Refolding Buffer | see JBC 271:21853 for detailed protocol |
| Pre-Refolding Purification | Reverse phase chromatography |
| Tag Cleaved | no tag |
| Refolding pH | 8.5 |
| Refolding Temperature | 4.0 |
| Protein Concentration | 0.25 mg/mL |
| Refolding Time | |
| Redox Agent | DTT |
| Redox Agent Concentration | n/a |
| Refolding Protocol | see JBC 271:21853 for detailed protocol |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 50% |
| Purity | |
| Notes | |