Small-molecule FRET probes for protein kinase activity monitoring in living cells

Vaasa, A., Lust, M., Terrin, A., Uri, A. and Zaccolo, M. (2010) Small-molecule FRET probes for protein kinase activity monitoring in living cells. Biochemical and Biophysical Research Communications, 397(4), pp. 750-755. (doi:10.1016/j.bbrc.2010.06.026)

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Abstract

In this study, the applicability of fluorescently labeled adenosine analogue-oligoarginine conjugates (ARC-Photo probes) for monitoring of protein kinase A (PKA) activity in living cells was demonstrated. ARC-Photo probes possessing subnanomolar affinity towards the catalytic subunit of PKA (PKAc) and competitive with the regulatory subunit (PKAr), penetrate cell plasma membrane and associate with PKAc fused with yellow fluorescent protein (PKAc-YFP). Detection of inter-molecular F÷rster resonance energy transfer (FRET) efficiency between the fluorophores of the fusion protein and ARC-Photo probe can be used for both the evaluation of non-labeled inhibitors of PKAc and for monitoring of cAMP signaling via detection of changes in the activity of PKA as a cAMP downstream effector.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Zaccolo, Professor Manuela and Terrin, Ms Anna
Authors: Vaasa, A., Lust, M., Terrin, A., Uri, A., and Zaccolo, M.
Subjects:Q Science > QH Natural history > QH345 Biochemistry
Q Science > QR Microbiology
College/School:College of Medical Veterinary and Life Sciences > Institute of Neuroscience and Psychology
Journal Name:Biochemical and Biophysical Research Communications
ISSN:0006-291X
ISSN (Online):1090-2104
Published Online:10 June 2010

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
479533Collaborative Research: Spatial and temporal aspects of cAMP/PKA signaling underlying information processing in neuronsManuela ZaccoloNational Institute of Health (NIH-GA)1R01AA 018060-0Institute of Neuroscience and Psychology
465341Role of PDE2 in the control of cardiac myocyte hypertrophy.Manuela ZaccoloBritish Heart Foundation (BHF)PG/07/091/23698Institute of Neuroscience and Psychology
432501Transatlantic networks of excellence in cardiovascular diseaseMiles HouslayFoundation Leducq (LEDUCQ-VIL)06 CVD 02Institute of Neuroscience and Psychology