Refolding Record:
| Protein | |
|---|---|
| Protein Name | Outer membrane protein A |
| Abbreviated Name | OmpA |
| SCOP Family | Outer membrane protein |
| Structure Notes | |
| Organism | Escherichia coli |
| UniProt Accession | P0A911 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Membrane and cell surface proteins and peptides |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 325 |
| Molecular Weight | 35172.3 |
| Pi | 5.59 |
| Molecular Weight | 35172.3 |
| Disulphides | 1 |
| Full Sequence |
APKDNTWYTGAKLGWSQYHDTGFINNNGPTHENQLGAGAFGGYQVNPYVGFEMGYDWLGRMPYKGSVENGAYKAQGVQLTAKLGYPITDDLDIYTRLGGMVWRADTKSNVYGKNHDTGVSPVFAGGVEYAITPEIATRLEYQWTNNIGDAHTIGTRPDNGMLSLGVSYRFGQGEAAPVVAPAPAPAPEVQTKHFTLKSDVLFNFNKATLKPEGQAALDQLYSQLSNLDPKDGSVVVLGYTDRIGSDAYNQGLSERRAQSVVDYLISKGIPADKISARGMGESNPVTGNTCDNVKQRAALIDCLAPDRRVEIEVKGIKDVVTQPQA
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Dornmair K, Kiefer H, Jähnig F. (1990) Biologycal Chemistry, 265, 18907-18911 |
| Project Aim | Protein refolding |
| Fusion | None |
| Protein Expression and Production | Protein expressed and purified in native conformation prior to denaturation and refolding. |
| Expression Host | None |
| Expression Strain | None |
| Expression Temp | 0.0 |
| Expression Time | 0 |
| Expression Vector | |
| Expression Protocol | |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | not specified |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Chaperone-assisted refolding |
| Wash Buffer | n/a |
| Solubilization Buffer | O.O5% SDS, 75 mM Tris/HCl buffer, pH = 7.5. |
| Refolding Buffer | Octylglucoside (OG) to a final concentration of 20 mg/ml, |
| Pre-Refolding Purification | not specified |
| Tag Cleaved | no tag |
| Refolding pH | 7.5 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | OmpA was dissolved at a concentration of 100 ug/ml in O.O5% SDS, 75 mM Tris/HCl buffer, pH = 7.5. This yielded a molar SDS to OmpA ratio of 500. Denaturation was achieved by boiling the sample for 10 min. Thereafter, the sample was put on ice for 2 min. Refolding was induced at room temperature by adding solid OG to a final concentration of 20 mg/ml, corresponding to a molar OG/SDS ratio of 40. |
| Refolding Assay | SDS-PAGE,Protease digestion |
| Refolding Chaperones | None |
| Refolding Additives | Octylglucoside (OG) |
| Additives Concentration | 20 mg/ml |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |