MRI biosensor for protein kinase A encoded by a single synthetic gene

Raag D. Airan, Amnon Bar-Shir, Guanshu Liu, Galit Pelled, Michael T Mcmahon, Peter C Van Zijl, Jeff W Bulte, Assaf A. Gilad

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1919-1923
Number of pages5
JournalMagnetic Resonance in Medicine
Volume68
Issue number6
DOIs
StatePublished - Dec 2012

Fingerprint

Synthetic Genes
Biosensing Techniques
Cyclic AMP-Dependent Protein Kinases
Protein Kinases
Phosphorylation
Magnetic Resonance Imaging
Tandem Repeat Sequences
Phosphotransferases
Peptides
Population
Genes

Keywords

  • biosensor
  • chemical exchange saturation transfer
  • protein kinase A
  • reporter gene

ASJC Scopus subject areas

  • Radiology Nuclear Medicine and imaging

Cite this

MRI biosensor for protein kinase A encoded by a single synthetic gene. / Airan, Raag D.; Bar-Shir, Amnon; Liu, Guanshu; Pelled, Galit; Mcmahon, Michael T; Van Zijl, Peter C; Bulte, Jeff W; Gilad, Assaf A.

In: Magnetic Resonance in Medicine, Vol. 68, No. 6, 12.2012, p. 1919-1923.

Research output: Contribution to journalArticle

Airan, Raag D. ; Bar-Shir, Amnon ; Liu, Guanshu ; Pelled, Galit ; Mcmahon, Michael T ; Van Zijl, Peter C ; Bulte, Jeff W ; Gilad, Assaf A. / MRI biosensor for protein kinase A encoded by a single synthetic gene. In: Magnetic Resonance in Medicine. 2012 ; Vol. 68, No. 6. pp. 1919-1923.
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