And if mutation has been done (in protein gene) and then there is no activity found with enzyme how one can monitor the function after mutation. Please explain both question if its possible.
You can find out if your protein is hexameric in solution by running it on a gelfiltration column preferentially coupled to a MALS setup to get a more accurate mass. If your solution mass corresponds to six times your monomeric mass you have a hexamer.
The second question is harder to answer... If there is functional assay you can try to monitor secondary structure by CD and solution behaviour by SEC. However this will only tell you if your molecule behaves differently in solution and if it remains folded and in the same oligomeric state not if your protein is actually active. One question that comes to mind is however why did you do mutations if you can not assess their effect? Or did you mean after the mutation there is no activity? Then you have your desired effect I suppose ... loss of function mutation.
- Badges
- Science method