Refolding Record:
| Protein | |
|---|---|
| Protein Name | Human leukocyte antigen DR3 beta chain |
| Abbreviated Name | HLA DR3 beta |
| SCOP Family | MHC antigen-recognition domain |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P01912 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha+Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | n |
| Domain | Extracellular domain only aa.1-198 |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 198 |
| Molecular Weight | 23052.6 |
| Pi | 6.48 |
| Molecular Weight | 23052.6 |
| Disulphides | 2 |
| Full Sequence |
G DTRPRFLEYS TSECHFFNGT ERVRYLDRYF HNQEENVRFD SDVGEFRAVT ELGRPDAEYW NSQKDLLEQK RGRVDNYCRH NYGVVESFTV QRRVHPKVTV YPSKTQPLQH HNLLVCSVSG FYPGSIEVRW FRNGQEEKTG VVSTGLIHNG DWTFQTLVML ETVPRSGEVY TCQVEHPSVT SPLTVEWRAR SESAQSK
|
| Notes | Transmembrane and cytoplasmic domains removed |
| Expression | |
|---|---|
| Report | Nag B, Arimilli S, Koukis B, Rhodes E, Baichwal V, Sharma SD (1994) J Biol Chem, 269, 10061-10070 |
| Project Aim | Structure-Function |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | None |
| Expression Temp | 37.0 |
| Expression Time | not stated |
| Expression Vector | not stated |
| Expression Protocol | not stated |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | Ion-exchange chromatography |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | n/a |
| Solubilization Buffer | 25mM phosphate pH 7.4, 8M urea, 20mM DTT |
| Refolding Buffer | phosphate buffered saline |
| Pre-Refolding Purification | Ion-exchange chromatography |
| Tag Cleaved | no tag |
| Refolding pH | 7.4 |
| Refolding Temperature | 4.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | DTT |
| Redox Agent Concentration | 20 |
| Refolding Protocol | E.coli inclusion bodies were solubilized in solubilization buffer. Initially the pellet was disruped by mild sonication, then it was allowed to sit at 37degC overnight. The suspension was centrifuged (1h, 100000g), the supernatant was then filtered. The protein was purified by ion exchange chromatography using a High Q-50 column, then refolded by dialysis against refolding buffer. |
| Refolding Assay | Online capillary isoelectric focussing electrospra,Isoelectic focusing |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | 95% |
| Notes | n/a |