Refolding Record:
| Protein | |
|---|---|
| Protein Name | Integration host factor beta subunit |
| Abbreviated Name | IHFb |
| SCOP Family | Prokaryotic DNA-bending protein |
| Structure Notes | |
| Organism | Escherichia coli |
| UniProt Accession | P0A6Y1 |
| 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 | 94 |
| Molecular Weight | 10651.1 |
| Pi | 9.34382 |
| Molecular Weight | 10651.1 |
| Disulphides | 0 |
| Full Sequence |
MTKSELIERL ATQQSHIPAK TVEDAVKEML EHMASTLAQG ERIEIRGFGS FSLHYRAPRT GRNPKTGDKV ELEGKYVPHF KPGKELRDRA NIYG
|
| Notes | subunit of the heterodimer IHF, partner protein is IHF-alpha subunit |
| Expression | |
|---|---|
| Report | Werner MH, Clore GM, Gronenborn AM, Kondoh A, Fisher RJ (1994) FEBS Letters, 345, 125-130 |
| 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 | HPLC |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Column refolding: size exclusion chromatography with partner subunits |
| Wash Buffer | n/a |
| Solubilization Buffer | 50mM TrisHCl, 200-500mM NaCl, 6-8M GdmCl pH 8.5 |
| Refolding Buffer | not specified |
| Pre-Refolding Purification | HPLC |
| Tag Cleaved | no tag |
| Refolding pH | 7.4 |
| Refolding Temperature | 25.0 |
| Protein Concentration | 1-2mM |
| Refolding Time | n/a |
| Redox Agent | None |
| Redox Agent Concentration | n/a,n/a,n/a |
| Refolding Protocol | The full protein complex was denatured in 8M GdmCl and the alpha and beta subunits were isolated and purified using reversed phase HPLC, then lyophilized. The subunits were then denatured in solubilization buffer, then refolded by passage down a gel filtration column with both the alpha and beta subunits, as well as a 30bp binding site oligonucleotide. |
| Refolding Assay | 15N -1H chemical shifts (ppm) |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | 60% |
| Purity | n/a |
| Notes | n/a |