target cytoplasmic protein to plasma membrane

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lotusgirl
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target cytoplasmic protein to plasma membrane

I would like to target a cytoplasmic protein to the plasma membrane. I want it to insert in the cell membrane and form a receptor with exposed domains on both sides. The protein is naturally capable of localising to the membrane by itself sometimes, but i want to enforce it's exptression at the cell membrane constitutively. I believe the way to do it is to add a signal sequence to import it into ER. Has anyone done this before? Are any references like published papers available where they have shown this a is a viable method? Need help with the theory as well as practical execution of the xept.

jenny99
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Quote:I want it to insert in

Quote:

I want it to insert in the cell membrane and form a receptor with exposed domains on both sides

you want to create a membrane protein that has an extracellular and an intracellular domain?

samm
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lotusgirl wrote:I would like

lotusgirl wrote:

I would like to target a cytoplasmic protein to the plasma membrane. I want it to insert in the cell membrane and form a receptor with exposed domains on both sides. The protein is naturally capable of localising to the membrane by itself sometimes, but i want to enforce it's exptression at the cell membrane constitutively. I believe the way to do it is to add a signal sequence to import it into ER. Has anyone done this before? Are any references like published papers available where they have shown this a is a viable method? Need help with the theory as well as practical execution of the xept.

Hi! This has definitely been done before - a Pubmed search turns up many references. However, just adding the signal sequence will most likely not be sufficient (though its the first thing to try - you might get lucky with your specific protein!). In order for your protein to express as a receptor, you need a membrane anchoring, and transmembrane domain - the sequences from CD16 and CD7, as well as other immunoglobulin motifs are fairly commonly used.
-sam