Date of Award
Spring 5-15-2023
Degree Name
Master of Science (MS)
Degree Type
Thesis
Abstract
Protein Kinase A (PKA) plays important roles in diverse biological processes such as sleep, long term memory, and synaptic plasticity. In addition, PKA also acts as an integrator of neuromodulator signaling though G protein-coupled receptor activation. However, despite genetic knockout and pharmacological inhibition experiments that demonstrate the importance of PKA, it is unclear where, when, or how PKA plays these roles in cellular physiology and behavior. In order to better understand the function of PKA in these processes, and how neuromodulator signaling drives complex behavioral changes, there exists a need for a method to selectively activate/inactivate PKA with high spatial and temporal precision. The photoreactive light, oxygen, or voltage 2 (LOV2) domain from Avena sativa phototropin 1 can be used to facilitate the activation/inactivation of an endogenous inhibitory peptide targeting the PKA catalytic subunit. The blue light induced conformational changes in LOV2 results in potent inhibition of PKA activity in the presence of blue light, and effective blocking of kinase-inhibitor interactions in the absence of light. We demonstrate the potency of this photoactivatable inhibitor peptide under dark and lit conditions, as well as the kinetics of caging/uncaging.
Language
English (en)
Chair
Yao Chen
Committee Members
Timothy Holy Jason Yi