Refolding Record:
| Protein | |
|---|---|
| Protein Name | Matrilysin |
| Abbreviated Name | MMP-7 |
| SCOP Family | Matrix metalloproteases, catalytic domain |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P09237 |
| 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 | 173 |
| Molecular Weight | 19130.5 |
| Pi | 8.7712 |
| Molecular Weight | 19130.5 |
| Disulphides | 0 |
| Full Sequence |
YSLFPN SPKWTSKVVT YRIVSYTRDL PHITVDRLVS KALNMWGKEI PLHFRKVVWG TADIMIGFAR GAHGDSYPFD GPGNTLAHAF APGTGLGGDA HFDEDERWTD GSSLGINFLY AATHELGHSL GMGHSSDPNA VMYPTYGNGD PQNFKLSQDD IKGIQKLYGK RSNSRKK
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Oneda H, Inouye K. (1999) J Biochem (Tokyo), 126, 905-911 |
| 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 | 37.0 |
| Expression Time | unknown |
| Expression Vector | unknown |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | soluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution/Column Refolding Combination |
| Wash Buffer | 2% Trition X-100 containing 10mM EDTA.2Na |
| Solubilization Buffer | 0.1M Tris-HCl containing 6M Gdn-HCl and 0.1M DTT |
| Refolding Buffer | 50mM Tris-HCl, 0.1mM Zn-acetate, 0.15M NaCl, 5mM CaCl2, 0.05% Brij-35, 10 mM oxidised GSH and 0.02% NaN3. |
| Pre-Refolding Purification | None |
| Tag Cleaved | yes |
| Refolding pH | 7.5 |
| Refolding Temperature | 4.0 |
| Protein Concentration | |
| Refolding Time | 12h |
| Redox Agent | DTT |
| Redox Agent Concentration | n/a |
| Refolding Protocol | unknown |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 0.32mg |
| Purity | |
| Notes | Refer to Kihira et al 1996 for method of expressing peptide |