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Review
. 2009 Oct;13(4):392-7.
doi: 10.1016/j.cbpa.2009.07.009. Epub 2009 Aug 12.

Fluorescent biosensors for real-time tracking of post-translational modification dynamics

Nwe-Nwe Aye-Han  1 , Qiang Ni, Jin Zhang
Affiliations
Review

Fluorescent biosensors for real-time tracking of post-translational modification dynamics

Nwe-Nwe Aye-Han et al. Curr Opin Chem Biol. 2009 Oct.

Abstract

Dynamic post-translational modifications (PTMs) regulate and diversify protein properties and cellular behaviors. Real-time monitoring of these modifications has been made possible with biosensors based on fluorescent proteins (FPs) and fluorescence resonance energy transfer (FRET), which can provide spatiotemporal information of PTMs with little perturbation to the cellular environment. In this review, we highlight available fluorescent biosensors applicable to detect PTMs in living cells and how they have shed light on biological questions that have been difficult to address otherwise. In addition, we also provide discussions about various engineering strategies for overcoming potential challenges associated with the development and application of such biosensors.

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Figures

Figure 1
Figure 1. General biosensor design and its application in living cells
(a) A schematic construction of unimolecular FRET-based biosensors for post-translational modification. The modular components are a FRET-donor and acceptor pair, a substrate domain that is subject to modification and a recognition domain that can recognize the particular post-translational modification. PTM dynamics can be measured by changes in FRET. (b) Temporal dynamics of PKA phosphorylation observed by PKA activity reporter, AKAR2. Black arrow indicates the activation of respective receptors.
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