Refolding Record:
| Protein | |
|---|---|
| Protein Name | Heparin-binding neurite-promoting factor |
| Abbreviated Name | HBNF |
| SCOP Family | Midkine, a heparin-binding growth factor, C-terminal domain |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P21246 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Small Proteins |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 137 |
| Molecular Weight | 15312.8 |
| Pi | 9.64 |
| Molecular Weight | 15312.8 |
| Disulphides | 5 |
| Full Sequence |
GKKEKPEKKVKKSDCGEWQWSVCVPTSGDCGLGTREGTRTGAECKQTMKTQRCKIPCNWKKQFGAECKYQFQAWGECDLNTALKTRTGSLKRALHNAECQKTVTISKPCGKLTKPKPQAESKKKKKEGKKQEKMLD
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Hulmes JD, Seddon AP, Decker MM, Böhlen P. (1993) Biochemical and Biophysical Research Com, 192, 738-746 |
| Project Aim | Structural Studies |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21(DE3)pLysS |
| Expression Temp | 0.0 |
| Expression Time | 00 |
| Expression Vector | pETHH8 |
| Expression Protocol | Human recombinant HBNF was expressed in E.coli BL@! pLYys containing the hHBNF-pETHH8 plasmid. |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | Not stated |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | n/a |
| Solubilization Buffer | 8 M urea, buffered at pH 8 containing 10 mM DTT. |
| Refolding Buffer | 25 mM HEPES (pH 7.4) |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.4 |
| Refolding Temperature | 0.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Inclusion bodies was solubuilised in 8 M urea, buffered at pH 8 containing 10 mM DTT. hrHBNF was refolded by dialysis againsed 25 mM HEPES (pH 7.4) and was purified by heparin affinity chromatography. |
| Refolding Assay | Mass spectrometry |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |