Please use this identifier to cite or link to this item:
Title: Insights into Protein Kinase A Activation using cAMP Analogs and Amide H/2H Exchange Mass Spectrometry
Keywords: Protein Kinase A, Amide Hydrogen Deuterium Exchange Mass Spectrometry, cAMP, Rp-cAMPS, RIα(91-244), PKA-C
Issue Date: 31-Jul-2009
Citation: BISHNOI TANUSHREE (2009-07-31). Insights into Protein Kinase A Activation using cAMP Analogs and Amide H/2H Exchange Mass Spectrometry. ScholarBank@NUS Repository.
Abstract: Cyclic adenosine 5?- monophosphate (cAMP) is an ancient signaling molecule and one of its primary eukaryotic targets is cAMP-dependent protein kinase A(PKA). PKA when inactive exists as a tetrameric complex of a dimeric regulatory subunit (PKA-R) and two monomeric catalytic subunits (PKA-C). The activity of PKA is regulated by binding of cAMP to the regulatory subunits in the inactive complex and releasing the PKA-C subunits. The signal for PKA-C dissociation is hypothesized to be propagated through two charge relays. The activation mechanism plays out through a ternary intermediate state consisting of the PKA-holoenzyme bound to cAMP, which occurs transiently before dissociation. The study of this transient complex can provide valuable insights into the activation mechanism of PKA and also help in the design of therapeutic molecules. This intermediate state was studied using Rp-cAMPS, a cAMP analog which is capable of locking the ternary complex. Rp-cAMPS blocks one of the two signal relays required for dissociation and allowed us to study the effects of the other. The changes observed upon binding Rp-cAMPS were localized on both the interacting subunits. The PKA-R subunit showed increased exchange in a region of the interaction interface. The PKA-C subunit also showed increased exchange in the corresponding interaction sites, which points at partial disruption of the interface between PKA-R and PKA-C subunits. Amide Hydrogen/Deuterium exchange followed by mass spectrometry was employed to compare the holoenzyme and the complex bound to Rp-cAMPS.
Appears in Collections:Master's Theses (Open)

Show full item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
TanushreeBishnoiThesis.pdf4.05 MBAdobe PDF



Page view(s)

checked on Apr 19, 2019


checked on Apr 19, 2019

Google ScholarTM


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.