Refolding Record:
| Protein | |
|---|---|
| Protein Name | Creatine kinase M-type |
| Abbreviated Name | MMCK |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Rabbit (Oryctolagus cuniculus) |
| UniProt Accession | P00563 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Dimer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 381 |
| Molecular Weight | 43112.1 |
| Pi | 6.63 |
| Molecular Weight | 43112.1 |
| Disulphides | Unknown |
| Full Sequence |
MPFGNTHNKYKLNYKSEEEYPDLSKHNNHMAKVLTPDLYKKLRDKETPSGFTLDDVIQTGVDNPGHPFIMTVGCVAGDEESYTVFKDLFDPIIQDRHGGFKPTDKHKTDLNHENLKGGDDLDPHYVLSSRVRTGRSIKGYTLPPHCSRGERRAVEKLSVEALNSLTGEFKGKYYPLKSMTEQEQQQLIDDHFLFDKPVSPLLLASGMARDWPDARGIWHNDNKSFLVWVNEEDHLRVISMEKGGNMKEVFRRFCVGLQKIEEIFKKAGHPFMWNEHLGYVLTCPSNLGTGLRGGVHVKLAHLSKHPKFEEILTRLRLQKRGTGGVDTAAVGSVFDISNADRLGSSEVEQVQLVVDGVKLMVEMEKKLEKGQSIDDMIPAQK
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Couthon F, Clottes E, Vial C. (1996) Biochem Biophys Res Commun., 227, 854-860 |
| 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 | None |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | 4.5 mM SDS-polyacrylamide, 100 mM Tris-acetate |
| Refolding Buffer | 100 mM Tris-acetate buffer containing OH-propyl B-cyclodextrin |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.4 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | 10 min |
| Redox Agent | None |
| Redox Agent Concentration | n/a,n/a,n/a |
| Refolding Protocol | Renaturation of SDS-denatured enzyme in the presence of hydroxy-propyl b-CD. MM-CK (25 mM),previously incubated for one hour in 4.5 mM SDS, was diluted ten-fold in 100 mM Tris-acetate pH 7.4 containing hydroxy-propyl -CD. A/ Reactivation as a function of time. |
| Refolding Assay | Fluorescence,Activity assay |
| Refolding Chaperones | None |
| Refolding Additives | β-cyclodextrin |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |