I want to test the activity of one kinase inside the cell. It is possible to purchase it as a recombinant protein. If I add it to the culture medium, will it get internalised by a mammalian cell to perform its function? Are there transporters on the cell membrane for kinase proteins?
Except for a few Tyrosine kinases, most kinases are intracellular proteins. And I doubt there are transporters for kinases. The best way to study kinase activity within the cell is to clone the gene into an expression construct, and transfect it into the cell and study its function. There are inducible constructs as well (like NAG repressible GlmS based ones) where you can switch on/off gene expression after transfection.
Hello
The cell membrane is selectively permeable allowing some substances to cross easily. Small molecules that are non-polar can cross the membrane through diffusion, but charged molecules and other molecules that are larger in size fail to cross the membrane.
The charged ions will enter the cell through protein channels embedded in the lipid bilayer of the membrane. Large molecules such as glucose will cross the membrane through diffusion.
So, for molecules to cross the cell membrane will need to resort to ion channels and specific transporters that require energy output from the cell.
In case of kinases, I have not come across any transporters for kinases. So, I doubt whether the kinase will get internalized by the mammalian cell. Moreover, there are other factors in the culture environment that could affect the activity of kinase. So, adding kinase to the culture medium would not be right.
Best.
If it does get across the membrane, it will be due to clearance receptors, such as LRP-1. From there it has a one-way ticket to lysosomes and degradation.
- Badges
- Science method
- Similar topics
- Mathematics
- Statistics
- Statistical Techniques
- Meta-Analysis