Dataset Information

Nanoscale regulation of Ca²⁺ dependent phase transitions and real-time dynamics of SAP97/hDLG.

ABSTRACT: Synapse associated protein-97/Human Disk Large (SAP97/hDLG) is a conserved, alternatively spliced, modular, scaffolding protein critical in regulating the molecular organization of cell-cell junctions in vertebrates. We confirm that the molecular determinants of first order phase transition of SAP97/hDLG is controlled by morpho-functional changes in its nanoscale organization. Furthermore, the nanoscale molecular signatures of these signalling islands and phase transitions are altered in response to changes in cytosolic Ca²⁺. Additionally, exchange kinetics of alternatively spliced isoforms of the intrinsically disordered region in SAP97/hDLG C-terminus shows differential sensitivities to Ca²⁺ bound Calmodulin, affirming that the molecular signatures of local phase transitions of SAP97/hDLG depends on their nanoscale heterogeneity and compositionality of isoforms.

SUBMITTER: Rajeev P

PROVIDER: S-EPMC9307800 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Nanoscale regulation of Ca<sup>2+</sup> dependent phase transitions and real-time dynamics of SAP97/hDLG.

Rajeev Premchand P Singh Nivedita N Kechkar Adel A Butler Corey C Ramanan Narendrakumar N Sibarita Jean-Baptiste JB Jose Mini M Nair Deepak D

Nature communications 20220722 1

Synapse associated protein-97/Human Disk Large (SAP97/hDLG) is a conserved, alternatively spliced, modular, scaffolding protein critical in regulating the molecular organization of cell-cell junctions in vertebrates. We confirm that the molecular determinants of first order phase transition of SAP97/hDLG is controlled by morpho-functional changes in its nanoscale organization. Furthermore, the nanoscale molecular signatures of these signalling islands and phase transitions are altered in respons ...[more]

PMID: 35869063

Similar Datasets

Project description:Adenosine 3',5'-cyclic monophosphate (cAMP) is a key second messenger that activates several signal transduction pathways in eukaryotic cells. Alteration of basal levels of cAMP is known to activate protein kinases, regulate phosphodiesterases and modulate the activity of ion channels such as Hyper polarization-activated cyclic nucleotide gated channels (HCN). Recent advances in optogenetics have resulted in the availability of novel genetically encoded molecules with the capability to alter cytoplasmic profiles of cAMP with unprecedented spatial and temporal precision. Using single molecule based super-resolution microscopy and different optogenetic modulators of cellular cAMP in both live and fixed cells, we illustrate a novel paradigm to report alteration in nanoscale confinement of ectopically expressed HCN channels. We characterized the efficacy of cAMP generation using ensemble photoactivation of different optogenetic modulators. Then we demonstrate that local modulation of cAMP alters the exchange of membrane bound HCN channels with its nanoenvironment. Additionally, using high density single particle tracking in combination with both acute and chronic optogenetic elevation of cAMP in the cytoplasm, we show that HCN channels are confined to sub 100 nm sized functional domains on the plasma membrane. The nanoscale properties of these domains along with the exchange kinetics of HCN channels in and out of these molecular zones are altered upon temporal changes in the cytoplasmic cAMP. Using HCN2 point mutants and a truncated construct of HCN2 with altered sensitivity to cAMP, we confirmed these alterations in lateral organization of HCN2 to be specific to cAMP binding. Thus, combining these advanced non-invasive paradigms, we report a cAMP dependent ensemble and single particle behavior of HCN channels mediated by its cyclic nucleotide binding domain, opening innovative ways to dissect biochemical pathways at the nanoscale and real-time in living cells.

Dataset Information

Nanoscale regulation of Ca²⁺ dependent phase transitions and real-time dynamics of SAP97/hDLG.

Publications

Nanoscale regulation of Ca<sup>2+</sup> dependent phase transitions and real-time dynamics of SAP97/hDLG.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Dataset Information

Nanoscale regulation of Ca2+ dependent phase transitions and real-time dynamics of SAP97/hDLG.

Publications

Nanoscale regulation of Ca<sup>2+</sup> dependent phase transitions and real-time dynamics of SAP97/hDLG.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Nanoscale regulation of Ca²⁺ dependent phase transitions and real-time dynamics of SAP97/hDLG.