Refolding Record:
| Protein | |
|---|---|
| Protein Name | ATP synthase epsilon chain |
| Abbreviated Name | ATPase epsilon |
| SCOP Family | Epsilon subunit of F1F0-ATP synthase C-terminal domain |
| Structure Notes | |
| Organism | Spinacia oleracea |
| UniProt Accession | P00833 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 136 |
| Molecular Weight | 14699.8 |
| Pi | 6.58883 |
| Molecular Weight | 14699.8 |
| Disulphides | 0 |
| Full Sequence |
MTLNLCVLTPNRSIWNSEVKEIILSTNSGQIGVLPNHAPTATAVDIGILRIRLNDQWLTL
ALMGGFARIGNNEITILVNDAERGSDIDPQEAQQTLEIAEANLRKAEGKRQKIEANLALR
RARTRVEASNTISS
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Steinemann D, Lill H, Junge W, Engelbrecht S. (1994) Biochim Biophys Acta., 1187, 354-359 |
| 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 |
| Expression Temp | 37.0 |
| Expression Time | up to 16h |
| Expression Vector | pJLA |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | Ion-exchange chromatography |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution/Column Refolding Combination |
| Wash Buffer | unknown |
| Solubilization Buffer | 8M urea, 50mM MES-NaOH pH 5.5 |
| Refolding Buffer | 50mM MES-NaOH, 500mM L-arginine |
| Pre-Refolding Purification | Ion-exchange chromatography |
| Tag Cleaved | no tag |
| Refolding pH | 7.5 |
| Refolding Temperature | 25.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Solubilized protein was purified to homogeneity on cation exchange chromatography. Fractions containing the target protein were initially diluted two-fold yielding a 4M urea solution. Subsequently the protein was fully refolded by gel filtration against 50mM MES-NaOH, 500mM L-arginine. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | homogeneous |
| Notes | |