Refolding Record:
| Protein | |
|---|---|
| Protein Name | Bone morphogenetic protein-2 |
| Abbreviated Name | BMP-2 |
| SCOP Family | Transforming growth factor (TGF)-beta |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P12643 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Small Proteins |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 116 |
| Molecular Weight | 12904.8 |
| Pi | 8.20959 |
| Molecular Weight | 12904.8 |
| Disulphides | 3 |
| Full Sequence |
QAKHKQRK RLKSSCKRHP
LYVDFSDVGW NDWIVAPPGY HAFYCHGECP FPLADHLNST NHAIVQTLVN SVNSKIPKAC
CVPTELSAIS MLYLDENEKV VLKNYQDMVV EGCGCR
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Vallejo LF, Brokelmann M, Marten S, Trappe S, Cabrera-Crespo J, Hoffmann A, Gross G, Weich HA, Rinas U. (2002) J Biotechnology, 94, 185-194 |
| Project Aim | Undefined |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21 |
| Expression Temp | 42.0 |
| Expression Time | 3h |
| Expression Vector | pCYT-EXP3 |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | High pressure homogenization |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | 20mM Tris-HCl, 0.5mM EDTA, 2% Triton X-100, pH 8.5 |
| Solubilization Buffer | 6M guanidinium chloride, 100mM Tris-HCl, 100mM DTT, 1mM EDTA, pH 8.5 |
| Refolding Buffer | 50mM Tris-HCl, 1M NaCl, 5mM EDTA, 1mM GSSG, 2mM GSH, 0.5M pyridine-3 sulfonic acid, 0.75M 2-(cyclohexylamino)ethanesulfonic acid, 1M 3-(1-pyridinio)-1-propanesulfonate |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.5 |
| Refolding Temperature | 20.0 |
| Protein Concentration | <0.1mg/ml |
| Refolding Time | 96h |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The inclusion bodies were washed vigourously by vortexing in 20mM Tris-HCl, 0.5mM EDTA, 2% Triton X-100, pH 8.5. The protein was subsequently solubilised in 6M guanidinium chloride, 100mM Tris-HCl, 100mM DTT, 1mM EDTA, pH 8.5 and refolded by dilution. The solution was left at 20C for 4 days. |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | >50% |
| Purity | 85% |
| Notes | |