Step 1: Determine which GPCRs are associated with the peach scent. (3.5 pts)
Insert the cDNA of all the olfactory GPCRs into E. coli and purify out all the olfactory GPCR proteins.
For each GPCR protein, have one control reaction with just the GPCR, and one experimental reaction where the GPCR is incubated with the peach extract. Do this for all the olfactory GPCRs.
Run all the samples on a gel. If a compound in the peach extract is the ligand for a specific GPCR, it will cause a conformational change in that GPCR. This will cause the band on the gel to shift relative to the control reaction for that GPCR.
This will allow us to identify which GPCRs are associated with the peach scent.
Step 2: Identify a single specific “peach” agonist that can stimulate all of the above determined GPCRs for ultimate incorporation in to a STINK™ Cartridge. Nintendo’s design specifications call for a single compound to be associated with each scent. (4 pts)
Find all the chemical compounds present in the peach extract using techniques such as spectroscopy
Gather the GPCR proteins found to be associated with peach scent, which were isolated in step 1
For each GPCR protein, have one control reaction with just the GPCR. Then several experimental reactions where the GPCR will be incubated with one of the compounds present in peach extract.
Run all of the samples on a gel. Analyze the results as described in step 1.
Look for a compound which causes a conformational change in all of the GPCRs associated with peach scent.
There are some drawbacks to Nintendo’s STINK™ delivery system. The machine can only deliver a small single size dose (a “puff”) of each compound. Despite this hardware flaw, Nintendo wishes to be able to control the extent and duration of each scent and they expect PBI to design methods by which to accomplish this fine control. Note that you can deliver a second “puff” of another compound. Consider how you might use each of the