Refolding Record:
| Protein | |
|---|---|
| Protein Name | Growth Hormone |
| Abbreviated Name | GH |
| SCOP Family | Long-Chain Cytokines |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P01241 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 191 |
| Molecular Weight | 22129.0 |
| Pi | 5.26 |
| Molecular Weight | 22129.0 |
| Disulphides | 2 |
| Full Sequence |
FPTI PLSRLFDNAM LRAHRLHQLA FDTYQEFEEA YIPKEQKYSF LQNPQTSLCF SESIPTPSNR EETQQKSNLE LLRISLLLIQ SWLEPVQFLR SVFANSLVYG ASDSNVYDLL KDLEEGIQTLMGRLEDGSPR TGQIFKQTYS KFDTNSHNDD ALLKNYGLLY CFRKDMDKVE TFLRIVQCRS VEGSCGF
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Yazdanparast R, Esmaeili MA, Khodagholi F. (2007) International J. Biological Macromolecul, 40, 126-133 |
| Project Aim | Protein refolding |
| Fusion | None |
| Protein Expression and Production | Protein expressed and purified in native conformation prior to denaturation and refolding. |
| Expression Host | None |
| Expression Strain | None |
| Expression Temp | 0.0 |
| Expression Time | 0 |
| Expression Vector | |
| Expression Protocol | |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | 100 mM Tris–base (pH 7) containing 4.5 M GuHCl |
| Solubilization Buffer | 100 mM Tris-buffer, 2mM CTAB |
| Refolding Buffer | 1.4 mM CTAB, 0.5 M GuHCl, 25 mM α-CD and/or 4.8 mM β-C |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | overnight |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Artificial chaperone-assisted refolding of denatured r-hGH in the presence of various α-CD (a) and β-CD (b) concentrations. The protein (2.7 mg/ml) was denatured for 4 h in 4.5 M GuHCl and then diluted with 100 mM Tris-buffer, pH 7, containing 2 mM CTAB. After 10 min incubation at room temperature, CD stock solutions was added to each sample to reach to the final concentrations of 0.3 mg/ml of r-hGH, 1.4 mM CTAB, 0.5 M GuHCl, 25 mM α-CD and/or 4.8 mM β-CD. The samples were incubated overnight at room temperature. The extent of aggregation was measured with respect to the respective control sample (devoid of detergent and cyclodextrin) and expressed as percent inhibition of aggregation. The error bars represent the standard error of mean for three independent measurements. Fig. 3c, represent the aggregation kinetics of denatured r-hGH in the presence of α- and/or β-CD as a function of time. |
| Refolding Assay | Fluorescence,Protein activity assay |
| Refolding Chaperones | None |
| Refolding Additives | β-cyclodextrin,α-cyclodextrin |
| Additives Concentration | 4.8 mM |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | In this experiment they compared different concentration of CTAB, TTAB,DTAB detergents in presence of a-CD and B-Cd in refolding. Results indicate that better refolding yield are obtained with CTAB in presence of a-CD |