Honors Theses

Date of Award

Spring 5-9-2020

Document Type

Undergraduate Thesis


Chemistry and Biochemistry

First Advisor

Nicole Ashpole

Second Advisor

Soumyajit Majumdar

Third Advisor

Jason Paris

Relational Format



Cannabinoid receptors CB1 and CB2 are G protein-coupled receptors that have a variety of physiological effects on the human body. Many natural product agonists and antagonists are presumed to interact with the cannabinoid receptors and are therefore heavily studied in drug discovery. Assays are used to study the binding of compounds to these receptors. However, a limited number of these assays are efficient in determining the functional activity of ligands associated with CB1 and CB2. Of those, many use radioactivity which is both expensive and potentially dangerous to the researcher. In this study, we first validated the assays already found to be effective in evaluating the function of ligands in the laboratory and then compared the results of the CB1 and CB2 assays. We found that although the cAMP-Glo and GTPγS assays were effective methods to measure functional efficacy for CB2, they were ineffective for CB1. Using compiled research methods from a number of academic journals, the establishment of a proper protocol for the functional characterization of ligands to CB1 was attempted but unsuccessful. To determine the possible reason for failure, a western blot was run to test the expression of G proteins in the inactive CB1 membrane in comparison to the G proteins in the active CB2 membrane. It was found that the G proteins in the active CB2 membrane were more heavily expressed than the G proteins in the inactive CB1 membrane. Together, these data concluded that the possible reason for our inability to establish an effective method to measure functional efficacy for CB1 is a lack of active G proteins in the membrane.



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