Refolding Record:
| Protein | |
|---|---|
| Protein Name | 18 kDa cyclophilin, Cyclophilin-18 |
| Abbreviated Name | C-18 |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Toxoplasma gondii |
| UniProt Accession | Q26994 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 163 |
| Molecular Weight | 17920.2 |
| Pi | 5.87 |
| Molecular Weight | 17920.2 |
| Disulphides | Unknown |
| Full Sequence |
ENAGVRKAYMDIDIDGEHAGRIILELREDIAPKTVKNFIGLFDKYKGSVFHRIIPDFMIQGGDFENHNGTGGHSIYGRRFDDENFDLKHERGVISMANAGPNTNGSQFFITTVKTEWLDARHVVFGKITTESWPTVQAIEALGGSGGRPSKVAKITDIGLLE
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Golding H, Khurana S, Yarovinsky F, King LR, Abdoulaeva G, Antonsson L, Owman C, Platt EJ, Kabat D, Andersen JF, Sher A. (2005) Biologycal Chemistry, 280, 29570-29577 |
| Project Aim | Undefined |
| 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 | 37.0 |
| Expression Time | 4 h |
| Expression Vector | n/a |
| Expression Protocol | Recombinant C-18 expression and purification steps were described elsewhere (15, 17). In brief, a plasmid encoding C-18 was transformed into Escherichia coli BL21(DE3)pLys (Invitrogen). Purified inclusion bodies obtained from bacteria induced with isopropyl-1-thio-B-D-galactopyranoside (Invitrogen) for 4 h were solubilized in 6 M guanidine HCl |
| Method of Induction | IPTG |
| 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 | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | 6 M guanidine HCl |
| Refolding Buffer | 20 mM Tris-HCl, pH 8.0, 150 mM NaCl buffer |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.0 |
| Refolding Temperature | 0.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The protein was refolded in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl buffer, dialyzed against 10 mM Tris-HCl, pH 8.0, buffer and purified on HiPrep 16/10 Q FF column (Amersham Biosciences), using 10 mM Tris-HCl, pH 8.0, 1 M NaCl for elution. |
| Refolding Assay | Unspecified |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |