Refolding Record:
| Protein | |
|---|---|
| Protein Name | Ribonuclease A |
| Abbreviated Name | RNase A |
| SCOP Family | Ribonuclease A-like |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P07998 |
| 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 | 128 |
| Molecular Weight | 14574.3 |
| Pi | 8.97 |
| Molecular Weight | 14574.3 |
| Disulphides | 4 |
| Full Sequence |
KE SRAKKFQRQH MDSDSSPSSS STYCNQMMRR RNMTQGRCKP VNTFVHEPLV DVQNVCFQEK VTCKNGQGNC YKSNSSMHIT DCRLTNGSRY PNCAYRTSPK ERHIIVACEG SPYVPVHFDA SVEDST
|
| Notes | A variety of mutants also constructed |
| Expression | |
|---|---|
| Report | Gaur D, Swaminathan S, Batra JK (2001) J Biol Chem, 276, 24978-24984 |
| Project Aim | Structure-Function,Biophysical Studies |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21(lambdaDE3) |
| Expression Temp | 37.0 |
| Expression Time | not stated |
| Expression Vector | pVEX11 |
| Expression Protocol | no details provided |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | Not stated |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | 6M GdmHCl |
| Refolding Buffer | contains L-arginine and GSSG |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.5 |
| Refolding Temperature | 4.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | GSSG |
| Redox Agent Concentration | n/a |
| Refolding Protocol | n/a |
| Refolding Assay | Far-UV Circular Dichroism,Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | L-Arginine |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |