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2015 ; 109
(6
): 1251-63
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Parallel Allostery by cAMP and PDE Coordinates Activation and Termination Phases
in cAMP Signaling
#MMPMID26276689
Krishnamurthy S
; Tulsian NK
; Chandramohan A
; Anand GS
Biophys J
2015[Sep]; 109
(6
): 1251-63
PMID26276689
show ga
The second messenger molecule cAMP regulates the activation phase of the cAMP
signaling pathway through high-affinity interactions with the cytosolic cAMP
receptor, the protein kinase A regulatory subunit (PKAR). Phosphodiesterases
(PDEs) are enzymes responsible for catalyzing hydrolysis of cAMP to 5' AMP. It
was recently shown that PDEs interact with PKAR to initiate the termination phase
of the cAMP signaling pathway. While the steps in the activation phase are well
understood, steps in the termination pathway are unknown. Specifically, the
binding and allosteric networks that regulate the dynamic interplay between PKAR,
PDE, and cAMP are unclear. In this study, PKAR and PDE from Dictyostelium
discoideum (RD and RegA, respectively) were used as a model system to monitor
complex formation in the presence and absence of cAMP. Amide hydrogen/deuterium
exchange mass spectrometry was used to monitor slow conformational transitions in
RD, using disordered regions as conformational probes. Our results reveal that RD
regulates its interactions with cAMP and RegA at distinct loci by undergoing slow
conformational transitions between two metastable states. In the presence of
cAMP, RD and RegA form a stable ternary complex, while in the absence of cAMP
they maintain transient interactions. RegA and cAMP each bind at orthogonal sites
on RD with resultant contrasting effects on its dynamics through parallel
allosteric relays at multiple important loci. RD thus serves as an integrative
node in cAMP termination by coordinating multiple allosteric relays and governing
the output signal response.