Please use this identifier to cite or link to this item:
Title: Defining the contours of cyclic nucleaotide mediated regulatory switches from prokaryotes to eukaryotes
Keywords: cAMP,Amide hydrogen/deuterium exchange,Cyclic nucleotide binding domain,Protein kinase A
Issue Date: 19-Aug-2011
Citation: SUGUNA BADIREDDY (2011-08-19). Defining the contours of cyclic nucleaotide mediated regulatory switches from prokaryotes to eukaryotes. ScholarBank@NUS Repository.
Abstract: cAMP signal transduction is a critical second messenger pathway in both prokaryotes and eukaryotes. We have used a combination of crystallography, amide hydrogen/deuterium exchange (HDX) and ion mobility mass spectrometry to study the cAMP mediated activation in eukaryotic PKA (Protein kinase A) as well as prokaryotic CNB (cyclic nucleotide binding domain) containing MSMEG_5458 and cynaobacterial GAF-B domain. Our study on isolated CNB-A of PKA provides evidence for conformational selection as the basis of cAMP action, contradicting earlier reports of induced fit mechanism alone. Furthermore, our study on PKA with tandem CNBs, reports that the close allosteric coupling between CNB-A and CNB-B with the C-subunit provides an important molecular insight into the function of CNB-B domain. Our HDX results of the MSMEG_5458 revealed a mechanism of cAMP mediated activation, in which binding of cAMP induces significant conformational changes that spatially rearranges the catalytic domain through a linker region, for activity. Our HDX results on GAF-B domain showed that isolated GAF-B bound to cAMP displayed an increased dynamics in comparison to cGMP, suggesting higher ordering of the protein in presence of cGMP. cAMP and cGMP induce distinct changes on the structural fold of the GAF-B domain. Extention of our analysis with GAF-AB has shown that GAF-AB lost its binding ability for cGMP. This suggests that the regulatory elements for ligand specificity are associated with GAF-A and the dimerization domain.
Appears in Collections:Ph.D Theses (Open)

Show full item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
Badireddy Suguna.pdf14.13 MBAdobe PDF



Page view(s)

checked on May 22, 2019


checked on May 22, 2019

Google ScholarTM


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