Refolding Record:
| Protein | |
|---|---|
| Protein Name | Flavohemoprotein (Globin domain) |
| Abbreviated Name | Flavohemoprotein |
| SCOP Family | Globins |
| Structure Notes | |
| Organism | Erwinia chrysanthemi |
| UniProt Accession | Q47266 |
| 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 | 401 |
| Molecular Weight | 44315.4 |
| Pi | 5.8874 |
| Molecular Weight | 44315.4 |
| Disulphides | 0 |
| Full Sequence |
MLDQQTIATIKSTIPLLAETGPALTAHFYQRMFHHNPELKDIFNMSNQRNGDQREALFNA
ICAYATHIENLPALLPAVERIAQKHASFNIQPEQYQIVGTHLLATLEEMFQPGQAVLDAW
GKRYGVLANVFIQRESDIYQQSAGQNGGWHGIRPFRIVAKQPQSSLITSFTLEPVDGGPI
AAFRPGQYLAVYIRDKRFEYQEIRQYSLTNEPNGRYYRIAVKRETMGSVSGYLHDVAREG
DVIELAAPHGDFYLEVTPETPVALISAGVGQTPMLSMLHSLKNQQHQADIFWLHAAENTE
VHAFADEIADVAATLPQLQSYVWYREASSEAARSAHAFHGLMALKDLPTPLPMTNLHCYL
CGPVAFMQFAARQLLELGITESQIHYECFGPHKVI
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Labesse G, Craescu CT, Mispelter J, Chottard G, Marden MC, Pin S, Forest E, Mornon JP, Boccara M. (1998) Eur J Biochem., 253, 751-759 |
| Project Aim | NMR |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21 |
| Expression Temp | 37.0 |
| Expression Time | 3-4h |
| Expression Vector | pT7.7 |
| Expression Protocol | unknown |
| 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 | Dilution |
| Wash Buffer | unknown |
| Solubilization Buffer | 6M urea |
| Refolding Buffer | 20mM sodium phosphate, 10mM KCN, 1.2-fold molar excess alkaline kemin-cyanide |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.5 |
| Refolding Temperature | 4.0 |
| Protein Concentration | |
| Refolding Time | 1h |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The solubilized inclusion bodies were refolded by slow 15-fold dilution in 20mM sodium phosphate, 10mM KCN, 1.2-fold molar excess alkaline kemin-cyanide pH 7.5 at 4 degrees C for 1h. The holoprotein was concentrated and purified on size exclusion chromatography. |
| Refolding Assay | 1H chemical shift (ppm) |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 100mg/L culture |
| Purity | |
| Notes | |