Refolding Record:
| Protein | |
|---|---|
| Protein Name | Pro-Falcipain-2 |
| Abbreviated Name | pro-Falcipain-2 |
| SCOP Family | Papain-like Cysteine Proteinases |
| Structure Notes | |
| Organism | Escherichia coli |
| UniProt Accession | Q9NBD4 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha+Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | |
| Domain | None |
| Chimera | None |
| Variants | None |
| Chain Length | 276 |
| Molecular Weight | 31419.6 |
| Pi | 5.52 |
| Molecular Weight | 31419.6 |
| Disulphides | Unknown |
| Full Sequence | |
| Notes | None |
| Expression | |
|---|---|
| Report | Kupper D., Reuter M., Mackeldanz P., Meisel A., Alves J., Schroeder C. and Kruger D. H. (1995) Protein Expression and Purification, 6, 1-9 |
| Project Aim | Crystallography |
| Fusion | N-terminal hexahistidine tag |
| 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 | 3 h |
| Expression Vector | pET3c |
| Expression Protocol | |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | Washing inclusion body |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | 50 mM TrisHCl (pH7.8), 10mM EDTA, 100mM NaCl, 0,5% Triton |
| Solubilization Buffer | 6M guanidine/HCL, 10 mM KH2PO4/K2HPO4 (pH 7.0), 1mM EDTA, 7mM ß-ME, 0,01% Triton |
| Refolding Buffer | 10 mM KH2PO4/K2HPO4 (pH 7.0), 1mM EDTA, 7mM ß-ME, 0,01% Triton |
| Pre-Refolding Purification | Washing inclusion body |
| Tag Cleaved | no |
| Refolding pH | 7.0 |
| Refolding Temperature | 4.0 |
| Protein Concentration | 0,1 µg/ml |
| Refolding Time | 24 h |
| Redox Agent | Beta-mercaptoethanol |
| Redox Agent Concentration | |
| Refolding Protocol | see: Protein Expr Purif. 1995 Feb;6(1):1-9. |
| Refolding Assay | enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 20 % |
| Purity | 99 % |
| Notes | |