Refolding Record:
| Protein | |
|---|---|
| Protein Name | Ribonuclease A |
| Abbreviated Name | RNase A |
| SCOP Family | Ribonuclease A-like |
| Structure Notes | |
| Organism | Bovine |
| UniProt Accession | P61823 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha+Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 126 |
| Molecular Weight | 13860.5 |
| Pi | 8.63544 |
| Molecular Weight | 13860.5 |
| Disulphides | 4 |
| Full Sequence |
LGKETA AAKFERQHMD SSTSAASSSN YCNQMMKSRN LTKDRCKPVN TFVHESLADV QAVCSQKNVA CKNGQTNCYQ SYSTMSITDC RETGSSKYPN CAYKTTQANK HIIVACEGNP YVPVHFDASV
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Werner MH, Clore GM, Gronenborn AM, Kondoh A, Fisher RJ (1994) FEBS Letters, 345, 125-130 |
| Project Aim | Undefined |
| 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 | |
| Expression Vector | |
| Expression Protocol | |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | not stated |
| Refolding | |
|---|---|
| Refolding Method | Column refolding: Size-exclusion chromatography |
| Wash Buffer | n/a |
| Solubilization Buffer | 50mM Tris-HCl, 50mM DTT, 200-500mM NaCl, 6-8M guanidinium chloride, pH 8.5 |
| Refolding Buffer | 20mM sodium phosphate, 200mM NaCl |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.5 |
| Refolding Temperature | 4.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | 1-10mg of protein was solubilized in 1-2ml of denaturation buffer, samples were allowed to stand several hours or overnight at ambient temperature. The protein was refolded on a Sephacryl S-100 column which was run at 4degC with a flow rate of 0.5ml/min. |
| Refolding Assay | 15N -1H chemical shifts (ppm),Unspecified |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | |
| Refolding Yield | |
| Purity | |
| Notes | |