Refolding Record:
| Protein | |
|---|---|
| Protein Name | Cell division topological specificity factor |
| Abbreviated Name | EcMinE |
| SCOP Family | Cell division protein MinE topological specificity domain |
| Structure Notes | |
| Organism | Escherichia coli |
| UniProt Accession | P0A734 |
| 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 | 88 |
| Molecular Weight | 10234.9 |
| Pi | 5.15 |
| Molecular Weight | 10234.9 |
| Disulphides | 0 |
| Full Sequence |
MALLDFFLSRKKNTANIAKERLQIIVAERRRSDAEPHYLPQLRKDILEVICKYVQIDPEMVTVQLEQKDGDISILELNVTLPEAEELK
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Aldridge, C., Moller, S.G. (2005) J Biol Chem, 280, 31673-31678 |
| Project Aim | Functional Studies |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21(D3) |
| Expression Temp | 37.0 |
| Expression Time | 2h |
| Expression Vector | pET14b |
| Expression Protocol | Cells were grown to a density of OD600:0.6 before induction with 1.5 mM ITPG. After incubation prote |
| Method of Induction | IPTG |
| Cell Density at Induction | OD 600 = 0.6 |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | Metal affinity chromatography |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | n/a |
| Solubilization Buffer | not stated |
| Refolding Buffer | 50 mM sodium phosphate, 50 mM NaCl, 0.1 M EDTA, 1.5 mM DTT, 10%glycerol, ph 7 over 8-0 urea gradient |
| Pre-Refolding Purification | Metal affinity chromatography |
| Tag Cleaved | no |
| Refolding pH | 7.0 |
| Refolding Temperature | 37.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | DTT |
| Redox Agent Concentration | 1.5 mM |
| Refolding Protocol | Refolding occured via dialysis against refolding buffer. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | Glycerol |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |