Refolding Record:
| Protein | |
|---|---|
| Protein Name | Geranylgeranylated Rab7 protein |
| Abbreviated Name | Rab7 |
| SCOP Family | G proteins |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P51149 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha/Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | n |
| Domain | n/a |
| Chimera | n/a |
| Variants | monoprenylated Rab7 |
| Chain Length | 207 |
| Molecular Weight | 23489.7 |
| Pi | 6.39 |
| Molecular Weight | 23489.7 |
| Disulphides | 0 |
| Full Sequence |
MTSRKKVLLKVIILGDSGVGKTSLMNQYVNKKFSNQYKATIGADFLTKEVMVDDRLVTMQ
IWDTAGQERFQSLGVAFYRGADCCVLVFDVTAPNTFKTLDSWRDEFLIQASPRDPENFPF
VVLGNKIDLENRQVATKRAQAWCYSKNNIPYFETSAKEAINVEQAFQTIARNALKQETEV
ELYNEFPEPIKLDKNDRAKASAESCSC
|
| Notes | Article did not clearly state which sequence the semisynthetic monoprenylated rab7 was derived from. |
| Expression | |
|---|---|
| Report | Alexandrov, K., Heinemann, I., Durek, T., Sidorovitch, V., Goody, R. S., Waldmann, H. (2002) J Am Chem Soc, 124, 5648-5649 |
| Project Aim | Structure-Function |
| Fusion | None |
| Protein Expression and Production | Protein synthesized by chemical means (not recombinant) and refolded. |
| Expression Host | None |
| Expression Strain | None |
| Expression Temp | 0.0 |
| Expression Time | n/a |
| Expression Vector | n/a |
| Expression Protocol | n/a |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Dilution/Dialysis combination |
| Wash Buffer | n/a |
| Solubilization Buffer | 100mM Tris-HCI, pH 8.0, 6M guanidinium hydrochloride, 1mM EDTA, 100mM DTE and 1% CHAPS |
| Refolding Buffer | 25mM Hepes, pH 7.5, 40mM NaCl, 2mM MgCl2, |
| Pre-Refolding Purification | None |
| Tag Cleaved | no |
| Refolding pH | 7.5 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The protein pellet was solubilized by addition of solubilizing buffer to final concentration of 0.2 mg/ml of ligated protein. This solution was diluted 20 fold with a buffer containing 100mM HEPES, pH 7.5, 5mM DTT, 5mM MgCl2, 20microM GDP, 5 mM DTE. After the addition of REP-1, to stabilise protein, the solution was dialyzed against the refolding buffer and concentrated using a pressure filtration unit. |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |