Refolding Record:
| Protein | |
|---|---|
| Protein Name | Gamma-crystallin C |
| Abbreviated Name | CRYGC |
| SCOP Family | C-type Lysozyme |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P07315 |
| 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 | 174 |
| Molecular Weight | 20874.7 |
| Pi | 6.88331 |
| Molecular Weight | 20874.7 |
| Disulphides | Unknown |
| Full Sequence |
MGKIPFYEDRA FQGRSYETTT DCPNLQPYFS RCNSIRVESG CWMLYERPNY QGQQYLLRRG EYPDYQQWMG LSDSIRSCCL IPQTVSHRLR LYEREDHKGL MMELSEDCPS IQDRFHLSEI RSLHVLEGCW VLYELPNYRG RQYLLRPQEY RRCQDWGAMD AKAGSLRRVV DLY
|
| Notes | Wild-type protein expressed solubly |
| Expression | |
|---|---|
| Report | Fu L, Liang JJN (2002) FEBS Letters, 513, 213-216 |
| Project Aim | Structural 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(DE3) |
| Expression Temp | 37.0 |
| Expression Time | 3h |
| Expression Vector | pET20b(+) |
| Expression Protocol | 1L LB media with 0.05mg/ml ampicillin was inoculated with a previously grown 20ml volume of cell culture. The cells were incubated at 37deg until OD590 reached 0.6-0.9, when expression was induced by the addition of 1mM IPTG. Incubation continued for another 3h when cells were harvested by centrifugation (10min x 5000rpm). |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD 590 = 0.6-0.9 |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | n/a |
| Solubilization Buffer | 4M GdnHCl |
| Refolding Buffer | not stated |
| Pre-Refolding Purification | None |
| Tag Cleaved | yes |
| Refolding pH | 7.5 |
| Refolding Temperature | 25.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | None |
| Redox Agent Concentration | n/a,n/a |
| Refolding Protocol | The inclusion bodies were solubilized in 4M Gdn and then refolded by dialysis. |
| Refolding Assay | Far-UV Circular Dichroism,Near-UV Circular Dichroism,Fluorescence,Western Blot,SDS-PAGE,Light scatter |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | |
| Refolding Yield | |
| Purity | |
| Notes | |