Refolding Record:
| Protein | |
|---|---|
| Protein Name | Methionine Adenosyltransferase |
| Abbreviated Name | MAT |
| SCOP Family | S-adenosylmethionine synthetase |
| Structure Notes | |
| Organism | Rat |
| UniProt Accession | P18298 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha+Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 401 |
| Molecular Weight | 43715.7 |
| Pi | 5.92521 |
| Molecular Weight | 43715.7 |
| Disulphides | 0 |
| Full Sequence |
MNGQLNGFHEAFIEEGTFLFTSESVGEGHPDKICDQINDAVLDAHLQQDPDAKVACETVA
KTGMILLAGEITSRAAIDYQKVVREAIKHIGYDDSSKGFDYKTCNVLVALEQQSPDIAQG
VHLDRNEEDIGAGDQGLMFGYATDETEECMPLTIVLAHKLNAKLAELRRNGTLPWLRPDS
KTQVTVQYMQDRGAVIPIRVHTIVISVQHDEEVCLDEMRDALKEKLIKAVVPAKYLDEDT
IYHLQPSGRFVIGGPQGDAGLTGRKIIVDTYGGWGAHGGGAFSGKDYTKVDRSAAYAARW
VAKSLVKGGLCRRVLVQVSYAIGVSHPLSISIFHYGTSQKSERELLEIVKNNFDLRPGVI
VRDLDLKKPIYQRTAAYGHFGRDSFPWEVPKKLKY
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Lopez-Vara MC, Gasset M, Pajares MA (2000) Protein Expression and Purification, 19, 219-226 |
| 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 | BL21 |
| Expression Temp | 37.0 |
| Expression Time | 3h |
| Expression Vector | pSSRL-T7N |
| 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 | soluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution/Dialysis combination |
| Wash Buffer | 100 mM Tris/HCl, pH 7.0, containing 5mMEDTA, 5 mMDTT, 4Murea, 5% Triton X-100, and 0.1 mM benzamidine |
| Solubilization Buffer | 50 mM Tris/HCl, pH 8, 8M urea, 75mM MgSO4 |
| Refolding Buffer | 50mM Tris-HCl, 10mM MgSO4, 10mM DTT |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.0 |
| Refolding Temperature | 4.0 |
| Protein Concentration | <0.1 mg/ml |
| Refolding Time | |
| Redox Agent | DTT |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Denaturant was initially diluted to <2M with 50mM Tris-HCl, pH 8.0. Subsequently, dialysis was performed against 50mM Tris-HCl, 10mM MgSO4, 10mM DTT, pH 8.0. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 88% |
| Purity | |
| Notes | |