Refolding Record:
| Protein | |
|---|---|
| Protein Name | Transforming growth factor beta 2 |
| Abbreviated Name | TGFbeta2 |
| SCOP Family | Transforming growth factor (TGF)-beta |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P61812 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Small Proteins |
| Molecularity | Dimer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 112 |
| Molecular Weight | 12719.6 |
| Pi | 7.68771 |
| Molecular Weight | 12719.6 |
| Disulphides | 5 |
| Full Sequence |
ALDAAYCF RNVQDNCCLR PLYIDFKRDL GWKWIHEPKG YNANFCAGAC PYLWSSDTQH SRVLSLYNTI NPEASASPCC VSQDLEPLTI LYYIGKTPKI EQLSNMIVKS CKCS
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Schlunegger MP, Cerletti N, Cox DA, McMaster GK, Schmitz A, Grutter MG (1992) FEBS Letters, 303, 91-93 |
| Project Aim | Crystallography |
| 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 | 5h |
| Expression Vector | pPLMu |
| Expression Protocol | Cells were grown at 30degC until OD550 reached 1.0, at which stage the temperature was shifted to 42degC and the incubation took place a further 5h. |
| Method of Induction | Temperature Shift |
| Cell Density at Induction | OD 550 = 1.0 |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | pH 2.5 |
| Refolding Buffer | redox system, CHAPS, pH 8.0 |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | Not specified |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Monomeric protein was solubized from inclusion bodies under acidic conditions and purified by size-exclusion chromatography. Refolding was performed in the presence of a redox system and a non-denaturing deteregent (CHAPS). Dimeric protein was then purified by cation-exchange chromatogaphy and reverse page HPLC. |
| Refolding Assay | Crystallography |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | |
| Refolding Yield | |
| Purity | |
| Notes | |