Refolding Record:
| Protein | |
|---|---|
| Protein Name | Retropepsin |
| Abbreviated Name | Retropepsin |
| SCOP Family | Retroviral protease (retropepsin) |
| Structure Notes | |
| Organism | HIV |
| UniProt Accession | P03369 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 99 |
| Molecular Weight | 10789.7 |
| Pi | 8.8308 |
| Molecular Weight | 10789.7 |
| Disulphides | 0 |
| Full Sequence |
PQITLWQRPL VTIRIGGQLK EALLDTGADD TVLEEMNLPG KWKPKMIGGI GGFIKVRQYD QIPVEICGHK AIGTVLVGPT PVNIIGRNLL TQIGCTLNF
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Cheng YS, McGowan MH, Kettner CA, Schloss JV, Erickson-Viitanen S, Yin FH. (1990) Gene, 87, 243-248 |
| Project Aim | Drug Studies |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | JM105 |
| Expression Temp | 37.0 |
| Expression Time | 3h |
| Expression Vector | pET3a |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | 5M urea |
| Solubilization Buffer | 8M urea, 50mM Tris-HCl, 1mM DTT, pH 8.0 |
| Refolding Buffer | 50mM Na-acetate, 10% glycerol, 1mM DTT, 0.1mM EDTA |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 5.5 |
| Refolding Temperature | 25.0 |
| Protein Concentration | 0.1-0.4 mg/ml |
| Refolding Time | 30min |
| Redox Agent | DTT |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The washed inclusion bodies were solubilized and diluted to 0.5-2mg/ml. The protein was refolded by 5-fold dilution in 50mM Na-acetate, 10% glycerol, 1mM DTT, 0.1mM EDTA pH 5.5 and was incubated for 30 min at 25 degrees C. The protein was purified to homogeneity by gel filtration. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | |
| Notes | |