NDLI: Visualization of PKA activity in plasma membrane microdomains.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Depry, Charlene Allen, Michael D. Zhang, Jin
Description	Country affiliation: United States Author Affiliation: Depry C ( Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.)
Abstract	Membrane rafts are sphingolipid- and cholesterol-rich microdomains that contain dynamic arrangements of signaling proteins. Notably, various components of the ubiquitous cAMP/Protein Kinase A (PKA) pathway, including ß-adrenergic receptors (ß-ARs), G proteins, and adenylyl cyclases (ACs), have been shown to localize differentially between membrane rafts and non-raft regions of the plasma membrane. As PKA participates in regulating diverse fundamental cellular functions, a number of which require membrane rafts, it is important to understand how PKA activity is specifically regulated in these membrane microdomains. To this end, we developed an improved FRET-based PKA activity biosensor, and targeted it to both membrane raft and non-raft regions of the plasma membrane to examine PKA activity dynamics in different plasma membrane microdomains. Disruption of membrane rafts via cholesterol depletion was shown to enhance ß-AR stimulated PKA activity at the plasma membrane, suggesting that membrane rafts play a negative role in ß-AR stimulated PKA activation. Furthermore, we found that membrane rafts possess higher basal PKA activity in the resting state compared to non-raft regions, which depends on the integrity of membrane rafts and proper localization of PKA. This study shows that membrane rafts play an important role in regulating the activity of PKA at the plasma membrane, and demonstrates the ability of live-cell FRET-based assays to reveal dynamic differences amongst plasma membrane microdomains, laying a foundation for further dissection of membrane regulated signal transduction.
File Format	HTM / HTML
ISSN	1742206X
Issue Number	1
Volume Number	7
e-ISSN	17422051
Journal	Molecular BioSystems
Language	English
Publisher	Royal Society of Chemistry
Publisher Date	2011-01-01
Publisher Place	Great Britain (UK)
Access Restriction	Subscribed
Subject Keyword	Discipline Molecular Biology Discipline Biochemistry Cell Membrane Enzymology Metabolism Cyclic Amp-dependent Protein Kinases Membrane Microdomains Biosensing Techniques Blotting, Western Cell Line Drug Effects Fluorescence Resonance Energy Transfer Humans Isoproterenol Pharmacology Receptors, Adrenergic, Beta Beta-cyclodextrins Journal Article Research Support, N.i.h., Extramural
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biotechnology

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