Refolding Record:
| Protein | |
|---|---|
| Protein Name | Beta crystallin A3 |
| Abbreviated Name | BA3 |
| SCOP Family | Crystallins/Ca-binding development proteins |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P05813 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Beta |
| Molecularity | Dimer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 235 |
| Molecular Weight | 27187.2 |
| Pi | 5.68 |
| Molecular Weight | 27187.2 |
| Disulphides | 0 |
| Full Sequence |
METQAEQQELETLPTTKMAQTNPTPGSLGPWKITIYDQENFQGKRMEFTSSCPNVSERSF
DNVRSLKVESGAWIGYEHTSFCGQQFILERGEYPRWDAWSGSNAYHIERLMSFRPICSAN
HKESKMTIFEKENFIGRQWEISDDYPSLQAMGWFNNEVGSMKIQSGAWVCYQYPGYRGYQ
YILECDHHGGDYKHWREWGSHAQTSQIQSIRRIQQDPAANKARKEAELAAATAEQ
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Bateman, O.A., Sarra, R., van Genesen, S.T., Kappe, G., Lubsen, N.H., Slingsby, C. (2003) Exp Eye Res, 77, 409-422 |
| Project Aim | Structural Studies |
| Fusion | C-terminal sequence |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21pLys |
| Expression Temp | 37.0 |
| Expression Time | 4 |
| Expression Vector | pET3a |
| Expression Protocol | 2L of LB containing ampicillin and chloramphenicol medium was inoculated with overnight culture. Cel |
| Method of Induction | IPTG |
| Cell Density at Induction | OD 600 = 0.3 |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | not stated |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | 9 M urea, 100mM NaPO4, pH 7.3 |
| Refolding Buffer | reduced concentrations of urea, 100mM NaPO4 |
| Pre-Refolding Purification | None |
| Tag Cleaved | no |
| Refolding pH | 7.3 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | overnig |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | BB1 was solubilized by addition of solubilization buffer and left overnight at room temperature. Refolding occured b the removal of urea. These solutions were also left overnight at room temperature. |
| Refolding Assay | Far-UV Circular Dichroism,Fluorescence |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |