Refolding Record:
| Protein | |
|---|---|
| Protein Name | Maltose-binding protein |
| Abbreviated Name | MalE or MBP |
| SCOP Family | Phosphate binding protein-like |
| Structure Notes | |
| Organism | Escherichia coli |
| UniProt Accession | P0AEX9 |
| 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 | 402 |
| Molecular Weight | 43387.6 |
| Pi | 5.53428 |
| Molecular Weight | 43387.6 |
| Disulphides | 0 |
| Full Sequence |
MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIK
VTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTW
DAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEP
YFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAE
AAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKE
LAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIP
QMSAFWYAVRTAVINAASGRQTVDEALKDAQTRITK
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Betton J, Hofnung M. (1996) J Biol Chem, 271, 8046-8052 |
| Project Aim | Undefined |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | K12 |
| Expression Temp | 37.0 |
| Expression Time | 3h |
| Expression Vector | pHCME |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | French Press |
| Lytic Agent | Lysozyme |
| Pre-Refolding Purification | None |
| Solubility | not stated |
| Refolding | |
|---|---|
| Refolding Method | Dilution/Dialysis combination |
| Wash Buffer | 10mM Tris-HCl Tris-HCl containing 0.7M sucrose and 1mM phenylsulfonyl fluroide. |
| Solubilization Buffer | 8M urea in 25mM Tris-HCl |
| Refolding Buffer | 2% Trition X-100 |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.5 |
| Refolding Temperature | 4.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Cells were harvested and suspended in 50 ml of 25 mM Tris-HCl buffer (pH 7.5) (buffer A) containing 0.1 mg/ml DNase I and lysed in a French press at 12,000 p.s.i. The lysate was centrifuged at 14,000 |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 23 micrograms per milligram |
| Purity | |
| Notes | |