Refolding Record:
| Protein | |
|---|---|
| Protein Name | Mitochondrial oxoglutarate-malate carrier protein |
| Abbreviated Name | OGC |
| SCOP Family | SLC25 |
| Structure Notes | |
| Organism | Bovine |
| UniProt Accession | P22292 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Membrane and cell surface proteins and peptides |
| Molecularity | Dimer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 318 |
| Molecular Weight | 34040.7 |
| Pi | 9.88749 |
| Molecular Weight | 34040.7 |
| Disulphides | 1 |
| Full Sequence |
AATASPGASGMDGKPRTSPKSVKFLFGGLAGMGATVFVQPLDLVKNRMQLSGEGAKTRE
YKTSFHALISILRAEGLRGIYTGLSAGLLRQATYTTTRLGIYTVLFERLTGADGTPPGFL
LKAVIGMTAGATGAFVGTPAEVALIRMTADGRLPVDQRRGYKNVFNALFRIVQEEGVPTL
WRGCIPTMARAVVVNAAQLASYSQSKQFLLDSGYFSDNILCHFCASMISGLVTTAASMPV
DIVKTRIQNMRMIDGKPEYKNGLDVLVKVVRYEGFFSLWKGFTPYYARLGPHTVLTFIFL
EQMNKAYKRLFLSG
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Fiermonte, G., Walker, J.E. and Palmieri, F. (1993) Biochem J., 294, 293-299 |
| 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(DE3) |
| Expression Temp | 37.0 |
| Expression Time | 5 h |
| Expression Vector | pKN172 |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | French Press |
| Lytic Agent | None |
| Pre-Refolding Purification | Washing inclusion body |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | none |
| Solubilization Buffer | 1.67% sarcosyl, 0.1 mM EDTA, 1 mM DTE and 10 mM Tris-HCl, pH 7.0 |
| Refolding Buffer | 0.6% sarcosyl, 0.03mM EDTA, 0.3 mM DTE and 3 mM Tris-HCl, pH 7.0 |
| Pre-Refolding Purification | Washing inclusion body |
| Tag Cleaved | no |
| Refolding pH | 7.0 |
| Refolding Temperature | 4.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | DTT |
| Redox Agent Concentration | n/a,n/a |
| Refolding Protocol | 1% of the inclusion bodies coming from 1 liter culture was solubilized in 60 ?l of the solubilization buffer consisting of 1.67% sarcosyl, 0.1 mM EDTA, 1 mM DTE (or DTT) and 10 mM Tris-HCl, pH 7.0. After 5 min in ice, the solubilization mixture was diluted 2.8 times with water (to reach a final concentration of detergent of 0.6%) and then centrifuged at 12000 g for 10 min at 4°C. 10 ?l of supernatant were reconstituted by the amberlite method (Palmieri, F., et al. Methods in Enzymol. 260, 349-369, 1995). |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | |
| Notes | |