Refolding Record:
| Protein | |
|---|---|
| Protein Name | Teratocarcinoma-derived growth factor 1 |
| Abbreviated Name | TDGF1 |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P13385 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | n |
| Domain | Epidermal growth factor-like domain aa.67-113 |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 47 |
| Molecular Weight | 5188.0 |
| Pi | 7.77 |
| Molecular Weight | 5188.0 |
| Disulphides | 3 |
| Full Sequence |
VPPMGIQHSKELNRTCCLNGGTCMLGSFCACPPSFYGRNCEHDVRKE
|
| Notes | Construct CR47C: 2nd and 4th N-terminal cys residues protected by acetamidomethyl (ACM) groups, all other cys residues triphenylmethyl protected |
| Expression | |
|---|---|
| Report | Lohmeyer M, Harrison PM, Kannan S, DeSantis M, OReilly NJ, Sternberg MJE, Salomon DS, Gullick WJ (1997) Biochemistry, 36, 3837-3845 |
| Project Aim | Structure-Function |
| Fusion | None |
| Protein Expression and Production | Protein synthesized by chemical means (not recombinant) and refolded. |
| Expression Host | None |
| Expression Strain | None |
| Expression Temp | 0.0 |
| Expression Time | n/a |
| Expression Vector | n/a |
| Expression Protocol | n/a |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | 6M urea, 0.05M glycine, 0.1M Tris pH 8.5 |
| Refolding Buffer | 20% DMSO, 1M urea, 0.05M glycine, 0.1M Tris pH 8.5 |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.5 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | 12h |
| Redox Agent | None |
| Redox Agent Concentration | n/a,n/a,n/a |
| Refolding Protocol | Peptides were synthesised, cleaved from the resin, partially deprotected and lyophilized. The peptide was then dissolved in deaerated helium-purged solubilization buffer, then sequentially dialysed againsed deaerated 4M and 2M urea buffers (0.05M glycine, 0.1M Tris pH 8.5). The protein was then refolded by 10-fold dilution against refolding buffer at room temperature for 12h with stirring. The peptide was then purified by preparative HPLC and lyophilized. The last two cysteines were deprotected and the the final formation of disulfide bondes was allowed in 1-ml of N2-purged H2O/MeOH (6:1 pH 3) and dropwise = addition of 5ml of 10mM I2 in MeOH followed by stirring for 40min at room temperature before quenching of the reaction. |
| Refolding Assay | Mass spectrometry |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |