help with experiment design - (Apr/16/2009 )

hi everyone
I need some help, I was asked to design & describe an experiment that will show the mechanism of the olfactory system in the molecular level with the knowledge that the olfactory receptors are probably from the G Protein coupled recptores (GPCRs) family and with the tools and technics that we have today.
I really need some ideas so please any one that can help is welcome

fadlone on Apr 16 2009, 12:27 AM said:

I would start by browsing a few papers by the nobel prize winners for the olfactory system (Richard Axel and Linda B. Buck, 2004) for inspiration.

fadlone on Apr 15 2009, 11:27 PM said:

You should isolate the cells that express those GPCRs with magnet bound antibodies against the GPCRs.
Then starting in the GPCR you should perform native far-western blot to detect the following interacting protein (1).
Then 1 with 2, 2 with 3.... This way you will find the proteins that follow the signal pathway starting in the GPCRs. Everytime you find the interaction you should caracterize the protein via MS, western blot or 2D electrophoresis. The last interacting protein should be one that interacts with DNA, to probe this interaction you can perform something called ChIP.

To probe the interaction between the olfactory molecule and GPCRs you can try to bind the olfactory molecule to a reporter: fluorescent, HRP...or make a rediactive isotope, but probably this demostration isn't worth the time and danger of working with radiactive isotopes.