a2 adrenergic receptor mutant uncoupled from Gi

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alexey-7
alexey-7's picture
a2 adrenergic receptor mutant uncoupled from Gi

Hello -

Is there a known mutant of a2(a) adrenergic receptor that can't bind Gi proteins?

I'm studying interactions between GPCRs and G proteins and it would be the best negative control for my experiments. What I need is a receptor that can't physically bind Gi/o proteins. I've done initial literature search but only found a2-adrenoceptor D79N mutant that is not able to modulate K+ currents but retains the ability to modulate Ca2+ and cAMP production.So,  I'm not sure how D79N mutation works and whether D79N can bind Gi proteins. If you know a better mutant please tell me.

Thank you
Alexey

Bluejay
Bluejay's picture
It would be much easier to

It would be much easier to try pertussis toxin.  100ng/mL overnight knocks out the A2A cell line we got from Perkin-Elmer.  Am not sure if this exactly answers your question.  (Both the calcium and label-free response were eliminated.)

alexey-7
alexey-7's picture
Thank you for your answer.

Thank you for your answer.

To make it clear I want to study precoupling between G proteins and GPCRs. So if I find any precoupling between Gi and a2-adrenoceptor then a2 adrenoceptor mutant that can not bind Gi would be a perfect negative control. Normally people use GPCRs that are not supposed to couple to a particular G protein or just use irrelevant membrane-tethered proteins as negative controls to exclude false positives due to non-specific interactions. However, I think the same receptor but mutated in a way that it does not couple to G protein anymore would be more appropriate. And that is a mutant I'm looking for.

Alexey