Refolding Record:
| Protein | |
|---|---|
| Protein Name | Human chorionic gonadotropin alpha subunit |
| Abbreviated Name | hCG-alpha |
| SCOP Family | Gonadotropin/Follitropin |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P01215 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Small Proteins |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 117 |
| Molecular Weight | 13075.2 |
| Pi | 8.53942 |
| Molecular Weight | 13075.2 |
| Disulphides | 5 |
| Full Sequence |
MDYYRKYAAIFLVTLSVFLHVLHSAPDVQDCPECTLQENPFFSQPGAPILQCMGCCFSRA
YPTPLRSKKTMLVQKNVTSESTCCVAKSYNRVTVMGGFKVENHTACHCSTCYYHKS
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Ren P, Sairam MR, Yarney TA. (1995) Mol Cell Endocrinol, 113, 39-51 |
| Project Aim | Functional Studies |
| Fusion | N-terminal hexahistidine tag |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | M15 |
| Expression Temp | 37.0 |
| Expression Time | 5h |
| Expression Vector | pQE-30 |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | Metal affinity chromatography |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | unknown |
| Solubilization Buffer | 6M guanidinium chloride, 0.1M NaH2PO4, 0.01M Tris-HCl, 10mM Imidazole, pH 8.0 |
| Refolding Buffer | Distilled water |
| Pre-Refolding Purification | Metal affinity chromatography |
| Tag Cleaved | no |
| Refolding pH | 6.1 |
| Refolding Temperature | 4.0 |
| Protein Concentration | |
| Refolding Time | 72h |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The whole cell pellet was harvested by centrifugation and solubilized in 6M guanidinium chloride, 0.1M NaH2PO4, 0.01M Tris-HCl, 10mM Imidazole, pH 8.0. After clarification by centrifugation the supernatant was purified on Ni-NTA, washing was performed with 8M urea, 0.1M NaH2PO4, 0.01M Tris-HCl, 10mM Imidazole, pH 6.1 and the protein was eluted in the same buffer containing 0.5M imidazole. Fraction containing the pure target protein were dialysed against distilled water for 3 days for refolding. |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | single band on SDS PAGE |
| Notes | |