Refolding Record:
| Protein | |
|---|---|
| Protein Name | Myotoxin alpha |
| Abbreviated Name | MTA |
| SCOP Family | Myotoxin |
| Structure Notes | |
| Organism | Crotalus viridis viridis |
| UniProt Accession | P01476 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Small Proteins |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 42 |
| Molecular Weight | 4787.7 |
| Pi | 9.47 |
| Molecular Weight | 4787.7 |
| Disulphides | 3 |
| Full Sequence |
YKQCHKKGGH CFPKEKICIG PSSDLGKMDC RWKWKCCKKG SG
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Nedelkov D, OKeefe MP, Chapman TL, Bieber AL (1997) Biochem Biophys Res Commun., 241, 252-259 |
| Project Aim | Structural Studies |
| Fusion | None |
| Protein Expression and Production | Protein synthesized by chemical means (not recombinant) and refolded. |
| Expression Host | None |
| Expression Strain | None |
| Expression Temp | 0.0 |
| Expression Time | 0 |
| 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 | |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | not stated |
| Refolding Buffer | 20mM phosphate pH 7.8, 0.2M GdmCl, 4.0mM GSH, 0.4mM GSSG |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.8 |
| Refolding Temperature | 25.0 |
| Protein Concentration | 7.5microM |
| Refolding Time | 12-16h |
| Redox Agent | GSH/GSSG |
| Redox Agent Concentration | 4.0/0.4mM |
| Refolding Protocol | The peptides were synthesised, then added to a final concentration of 7.5microM in refolding buffer. The solution (1000ml) final volume was capped with air, shaken vigorously and left at room temperature overnight. The protein was then purified by cation exchange chromatography |
| Refolding Assay | HPLC,Mass spectrometry |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | 5-7% |
| Purity | n/a |
| Notes | n/a |