Refolding Record:
| Protein | |
|---|---|
| Protein Name | ProCathepsin D |
| Abbreviated Name | ProCatD |
| SCOP Family | Pepsin-like proteases |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P07339 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 418 |
| Molecular Weight | 44552.2 |
| Pi | 6.10412 |
| Molecular Weight | 44552.2 |
| Disulphides | 4 |
| Full Sequence |
MQPSSLLPLALCLLAAPASALVRIPLHKFTSIRRTMSEVGGSVEDLIAKGPVSKYSQAVP
AVTEGPIPEVLKNYMDAQYYGEIGIGTPPQCFTVVFDTGSSNLWVPSIHCKLLDIACWIH
HKYNSDKSSTYVKNGTSFDIHYGSGSLSGYLSQDTVSVPCQSASSASALGGVKVERQVFG
EATKQPGITFIAAKFDGILGMAYPRISVNNVLPVFDNLMQQKLVDQNIFSFYLSRDPDAQ
PGGELMLGGTDSKYYKGSLSYLNVTRKAYWQVHLDQVEVASGLTLCKEGCEAIVDTGTSL
MVGPVDEVRELQKAIGAVPLIQGEYMIPCEKVSTLPAITLKLGGKGYKLSPEDYTLKVSQ
AGKTLCLSGFMGMDIPPPSGPLWILGDVFIGRYYTVFDRDNNRVGFAEAARL
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Sachdev D, Chirgwin JM. (1998) Protein Expression and Purification, 12, 122-132 |
| Project Aim | Undefined |
| Fusion | C-terminal maltose binding protein |
| 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 | 2-3h |
| Expression Vector | pET23b |
| 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 | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | 50mM Tris-HCl, 5mM EDTA, 0.02% TWEEN 80, pH 8.0 |
| Solubilization Buffer | 8M urea, 0.1M phosphate, 5mM EDTA, pH 7.0 |
| Refolding Buffer | 0.1M phosphate, 5mM EDTA, 5mM cysteine |
| Pre-Refolding Purification | None |
| Tag Cleaved | yes |
| Refolding pH | 7.0 |
| Refolding Temperature | 4.0 |
| Protein Concentration | |
| Refolding Time | 15h |
| Redox Agent | Cysteine |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Protein folds and remains soluble, however it is not active as a protease. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | |
| Notes | |