Refolding Record:
| Protein | |
|---|---|
| Protein Name | Toxin 5 |
| Abbreviated Name | Cn5 |
| SCOP Family | Long-chain scorpion toxins |
| Structure Notes | |
| Organism | Scorpion (C. noxius) |
| UniProt Accession | P45663 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Small Proteins |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 88 |
| Molecular Weight | 9480.1 |
| Pi | 8.57285 |
| Molecular Weight | 9480.1 |
| Disulphides | 4 |
| Full Sequence |
MNSLLMITACLFLIGTVWAKEGYLVNKSTGCKYGCLLLGKNEGCDKECKAKNQGGSYGYC
YAFGCWCEGLPESTPTYPLPNKSCSKK
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Altamirano MM, Garcia C, Possani LD, Fersht AR. (1999) Nat Biotechnol., 17, 187-191 |
| Project Aim | Structural Studies |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | None |
| Expression Temp | 37.0 |
| Expression Time | unknown |
| Expression Vector | unknown |
| 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 | soluble |
| Refolding | |
|---|---|
| Refolding Method | Column refolding: Oxidative refolding chromatography |
| Wash Buffer | 100 l of 6 M guanidinium chloride prepared in 100 mM potassium phosphate buffer (pH 8) |
| Solubilization Buffer | 6M guandinium chloride in 100mM phosphate buffer |
| Refolding Buffer | 100 mM potassium phosphate at pH 8, 0.5 M L?arginine, 1 mM GSSG (glutathione oxidized form), 1 mM GSH (glutathione reduced form) and 2 mM sodium EDTA. |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | 2.50 mg/ml |
| Refolding Time | 4 |
| Redox Agent | DTT |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Refer to: http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=9114491 |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 98% |
| Purity | |
| Notes | |