MRI biosensor for protein kinase A encoded by a single synthetic gene
Purpose: Protein kinases including protein kinase A (PKA) underlie myriad important signaling pathways. The ability to monitor kinase activity in vivo and in real‐time with high spatial resolution in genetically specified cellular populations is a yet unmet need, crucial for understanding complex bi...
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Published in | Magnetic resonance in medicine Vol. 68; no. 6; pp. 1919 - 1923 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Hoboken
Wiley Subscription Services, Inc., A Wiley Company
01.12.2012
Wiley Subscription Services, Inc |
Subjects | |
Online Access | Get full text |
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Summary: | Purpose:
Protein kinases including protein kinase A (PKA) underlie myriad important signaling pathways. The ability to monitor kinase activity in vivo and in real‐time with high spatial resolution in genetically specified cellular populations is a yet unmet need, crucial for understanding complex biological systems as well as for preclinical development and screening of novel therapeutics.
Methods:
Using the hypothesis that the natural recognition sequences of protein kinases may be detected using chemical exchange saturation transfer magnetic resonance imaging, we designed a genetically encoded biosensor composed of eight tandem repeats of the peptide LRRASLG, a natural target of PKA.
Results:
This sensor displays a measurable change in chemical exchange saturation transfer signal following phosphorylation by PKA. The natural PKA substrate LRRASLG exhibits a chemical exchange saturation transfer‐magnetic resonance imaging contrast at +1.8 and +3.6 ppm, with a >50% change after phosphorylation with minutes‐scale temporal resolution. Expression of a synthetic gene encoding eight monomers of LRRASLG yielded two peaks at these chemical exchange saturation transfer frequencies.
Conclusion:
Taken together, these results suggest that this gene may be used to assay PKA levels in a biologically relevant system. Importantly, the design strategy used for this specific sensor may be adapted for a host of clinically interesting protein kinases. Magn Reson Med, 2012. © 2012 Wiley Periodicals, Inc. |
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Bibliography: | NIH - No. EB008769; No. NS065284; No. EB005252; No. EB012590; No. EB006394; No. NS072171; No. EB015032 ArticleID:MRM24483 istex:7E7A0DA8F83ABDEA97B8D9611EA604646E871053 ark:/67375/WNG-S6B13C8D-M ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0740-3194 1522-2594 |
DOI: | 10.1002/mrm.24483 |