1 reply to this topic

[fadlone]

hello everyone

I need some suggestion on how to check activity of GPCR when a ligand bind to it, some one suggest me to use GFP gene reporter but I didn't quite understood how it will help
do if you get any ideas please help

thanks

[Amy831]

If there is a change in enzyme activity (downstream of the receptor) or second messenger production upon ligand binding and GPCR activation, and the signaling pathway(s) is known for your GPCR, you can use these parameters as a measure of activity.

As examples, adenylate cyclase (AC) is activated by some GPCRs, and you could look at enzyme activity with an assay specific for this enzyme, or an enzyme in your GPCR's pathway.

An alternative example would be to look at cAMP production if this is a downstream messenger for your GPCR. There are some newer, non-radioactive 96-well format assays to measure cAMP synthesis. Otherwise, there are similar assays for other second messengers.

fadlone, on Apr 23 2009, 01:31 PM, said:

hello everyone

I need some suggestion on how to check activity of GPCR when a ligand bind to it, some one suggest me to use GFP gene reporter but I didn't quite understood how it will help
do if you get any ideas please help

thanks