Refolding Record:
| Protein | |
|---|---|
| Protein Name | Outer membrane protein OprM |
| Abbreviated Name | OprM |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Pseudomonas aeruginosa |
| UniProt Accession | Q51487 |
| 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 | 493 |
| Molecular Weight | 52598.2 |
| Pi | 5.52071 |
| Molecular Weight | 52598.2 |
| Disulphides | Unknown |
| Full Sequence |
MKRSFLSLAVAAVVLSGCSLIPDYQRPEAPVAAAYPQGQAYGQNTGAAAVPAADIGWREF
FRDPQLQQLIGVALENNRDLRVAALNVEAFRAQYRIQRADLFPRIGVDGSGTRQRLPGDL
STTGSPAISSQYGVTLGTTAWELDLFGRLRSLRDQALEQYLATEQAQRSAQTTLVASVAT
AYLTLKADQAQLQLTKDTLGTYQKSFDLTQRSYDVGVASALDLRQAQTAVEGARATLAQY
TRLVAQDQNALVLLLGSGIPANLPQGLGLDQTLLTEVPAGLPSDLLQRRPDILEAEHQLM
AANASIGAARAAFFPSISLTANAGTMSRQLSGLFDAGSGSWLFQPSINLPIFTAGSLRAS
LDYAKIQKDINVAQYEKAIQTAFQEVADGLAARGTFTEQLQAQRDLVKASDEYYQLADKR
YRTGVDNYLTLLDAQRSLFTAQQQLITDRLNQLTSEVNLYKALGGGWNQQTVTQQQTAKK
EDPQA
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Charbonnier F, Kohler T, Pechere JC, Ducruix A. (2001) Protein Expression and Purification, 23, 121-127 |
| Project Aim | Functional Studies |
| 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 |
| Expression Temp | 37.0 |
| Expression Time | 3 hours |
| Expression Vector | pET28a/pET28b |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | Lysozyme |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution/Column Refolding Combination |
| Wash Buffer | 20 mM KH(2)PO(4) |
| Solubilization Buffer | 6M Guanidinium Hydrochloride, 1% (w/v) n-octylpolyoxyethylene, 2% (w/v) Triton X-100 |
| Refolding Buffer | 20-500 mM Imidazole gradient, 0.6% (w/v) C8E4. |
| Pre-Refolding Purification | None |
| Tag Cleaved | yes |
| Refolding pH | 8.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | Beta-mercaptoethanol |
| Redox Agent Concentration | n/a |
| Refolding Protocol | After growing at 37 |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 6 mg per litre cell culture |
| Purity | 80% |
| Notes | |