Refolding Record:
| Protein | |
|---|---|
| Protein Name | H-ras |
| Abbreviated Name | H-ras |
| SCOP Family | G proteins |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P01112 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha/Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 192 |
| Molecular Weight | 21298.1 |
| Pi | 5.15723 |
| Molecular Weight | 21298.1 |
| Disulphides | 0 |
| Full Sequence |
MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVIDGETCLLDILDTAG
QEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVKDSDDVPMVLVGNKCDL
AARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREIRQHKLRKLNPPDESGPG
CMSCKCVLS
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | DeLoskey RJ, Van Dyk DE, Van Aken TE, Campbell-Burk S. (1994) Archives Biochemistry and Biophysics, 311, 72-78 |
| Project Aim | Undefined |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | JM105 |
| Expression Temp | 30.0 |
| Expression Time | 4h |
| Expression Vector | pTAC |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | partial |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | unknown |
| Solubilization Buffer | 5M guanidinium chloride, 50mM Tris-HCl, 50mM NaCl, 5mM MgCl2, 1mM EDTA, 5mM DTT, 1mM PMSF, 30micromolar GDP, 10% glycerol, pH 8.0 |
| Refolding Buffer | 50mM Tris-HCl, 50mM NaCl, 5mM MgCl2, 1mM EDTA, 1mM DTT, 1mM PMSF, 30micromolar GDP, 10% glycerol |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.0 |
| Refolding Temperature | 4.0 |
| Protein Concentration | less than 0.1mg/ml |
| Refolding Time | 2h |
| Redox Agent | DTT |
| Redox Agent Concentration | n/a |
| Refolding Protocol | unknown |
| Refolding Assay | Ligand Binding |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 50-80% |
| Purity | |
| Notes | |