In:
Magnetic Resonance in Medicine, Wiley, Vol. 68, No. 6 ( 2012-12), p. 1919-1923
Kurzfassung:
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.
Materialart:
Online-Ressource
ISSN:
0740-3194
,
1522-2594
Sprache:
Englisch
Verlag:
Wiley
Publikationsdatum:
2012
ZDB Id:
1493786-4
