Refolding Record:
| Protein | |
|---|---|
| Protein Name | C1-Inhibitor |
| Abbreviated Name | C1-Inhibitor |
| SCOP Family | Serpins |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P05155 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Multi-domain proteins (alpha and beta) |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 478 |
| Molecular Weight | 52843.4 |
| Pi | 5.97 |
| Molecular Weight | 52843.4 |
| Disulphides | 2 |
| Full Sequence |
NPNATSSS SQDPESLQDR GEGKVATTVI SKMLFVEPIL EVSSLPTTNS TTNSATKITA NTTDEPTTQP TTEPTTQPTI QPTQPTTQLP TDSPTQPTTG SFCPGPVTLC SDLESHSTEA VLGDALVDFS LKLYHAFSAM KKVETNMAFS PFSIASLLTQ VLLGAGENTK TNLESILSYP KDFTCVHQAL KGFTTKGVTS VSQIFHSPDL AIRDTFVNAS RTLYSSSPRV LSNNSDANLE LINTWVAKNT NNKISRLLDS LPSDTRLVLL NAIYLSAKWK TTFDPKKTRM EPFHFKNSVI KVPMMNSKKY PVAHFIDQTL KAKVGQLQLS HNLSLVILVP QNLKHRLEDM EQALSPSVFK AIMEKLEMSK FQPTLLTLPR IKVTTSQDML SIMEKLEFFD FSYDLNLCGL TEDPDLQVSA MQHQTVLELT ETGVEAAAAS AISVARTLLV FEVQQPFLFV LWDQQHKFPV FMGRVYDPRA
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Gauthier M, Patston PA. (1997) Analytical Biochemistry, 248, 228-233 |
| Project Aim | Undefined |
| Fusion | None |
| Protein Expression and Production | Protein expressed and purified in native conformation prior to denaturation and refolding. |
| Expression Host | Escherichia coli |
| Expression Strain | None |
| Expression Temp | 37.0 |
| Expression Time | |
| Expression Vector | |
| Expression Protocol | |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Column refolding: Size-exclusion chromatography |
| Wash Buffer | n/a |
| Solubilization Buffer | 6M guanidinium chloride |
| Refolding Buffer | 8.1mM Na2HPO4, 1.5mM KH2PO4, 140mM NaCl, 2.7mM Kcl, 0.02% NaN3 |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.4 |
| Refolding Temperature | 25.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | This refolding study was carried out using pre-purified polymeric C1-inhibitor, and not inclusions bodies. C1-inhibitor polymers were prepared by heating the protein at 60C for 5 min at 10mg/ml. 0.2ml samples of unfolded protein at 5mg/ml were applied to gel filtration and run at 0.5ml/min. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 47% |
| Purity | |
| Notes | |