Refolding Record:
| Protein | |
|---|---|
| Protein Name | Type 1 ribosome-inactivating protein musarmin le |
| Abbreviated Name | MU1 |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Muscari armeniacum L. |
| UniProt Accession | Q8LLV7 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 327 |
| Molecular Weight | 36225.6 |
| Pi | 7.07 |
| Molecular Weight | 36225.6 |
| Disulphides | 0 |
| Full Sequence |
MKYLLPTAAAGLLLLAAQPAMAMAGQGFLTVEFTKTLNAVTLTSSTYTTFIRELRSRLAQTYVAGVPNIPVLPVYNQTQPPQGFDIVLTAGADRTTVRFRRDTLYLVGYQMQSGAWLEFGRAGNPQFIRGSEFLGFGGSYTELERLAGPVTSMDINQAKLVTSVRDLAVSTNSVVRAKALVVLIQMICEATRFIPISDHFASNLATDYAKLPPWMMSDLEKNWARISREVLKWDADTSYKIQPQTINGQTITTVEGLRPYLGILYRASVNPFSTSLYEEMKVGLEFAARRFGPVMVALAAALEIKRASQPELAPEDPEDVEHHHHHH
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Antolin, P., Girotti, A., Arias, F. S., Barriuso, B., Jimenez, P., Rojo, M. A., Girbes, T. (2004) J Biotechnology, 112, 313-322 |
| Project Aim | Structure-Function,Recombinant Protein Expression |
| Fusion | C-terminal hexahistidine tag |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21(DE3)pLysS |
| Expression Temp | 25.0 |
| Expression Time | 6h |
| Expression Vector | pET-25b(+) |
| Expression Protocol | Cells grown at 25degC in LB medium contqaining ampicillin and chloramphenicol. They were then induce |
| Method of Induction | IPTG |
| Cell Density at Induction | OD 600 = 0.8 |
| Cell Disruption Method | Freeze-thaw |
| Lytic Agent | Lysozyme |
| Pre-Refolding Purification | Washing inclusion body |
| Solubility | partial |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | n/a |
| Solubilization Buffer | 1.5% (w/v) cetyltrimethylammonium bromide |
| Refolding Buffer | 5 mM K2HPO4, pH 8.0 |
| Pre-Refolding Purification | Washing inclusion body |
| Tag Cleaved | no |
| Refolding pH | 8.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Inclusion bodies washed several times and suspended in solubilization buffer. After removing insoluble material by centrifugation solution dialysed against refolding buffer to obtain biologically active protein. |
| Refolding Assay | Enzyme activity,Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | Glycerol |
| Additives Concentration | 20% |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |