Cyclic adenosine monophosphate (cAMP) is an important, ubiquitous second messenger in cells. Phosphodiesterase (PDE) is the critical family of enzymes that hydrolyze cAMP in order to modulate cAMP signaling. cAMP concentrations are thought to be compartmentalized, that is, unique at different locations throughout a cell. Because PDE reacts with specific local concentrations of cAMP within each compartment, it may be possible to estimate the concentration of a known type of PDE in a specific area of the cell. This project is focused on determining the concentration of PDE in the near plasma membrane via cAMP measurements using cyclic nucleotide-gated CNG ion channels and computer modeling techniques. The activity of CNG channels was used to estimate cAMP levels near the plasma membrane. Modeling techniques were implemented in order to estimate subcellular PDE concentrations based upon the time course and amplitude of measured cAMP signals. Results indicate that that PDE levels near the plasma membrane are substantially lower than in the rest of the cell. These results will help us to better understand how signaling specificity is achieved within the cAMP pathway.

Estimating the near-membrane concentration of phosphodiesterase in HEK-293 cells

247th National Meeting (2014)

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