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Sensing second messengers

Fluorescent protein-based biosensors

Nature Cell Biology volume 11, pages S20–S21 (2009)Cite this article

These two Ca2+ biosensors were the first genetically encoded molecular sensors.

Ca2+ binds to calmodulin (CaM), changing its conformation and allowing it to bind to target proteins. Miyawaki et al. made a construct encoding blue GFP, CaM, the CaM-binding peptide of myosin light-chain kinase (M13) and green GFP. On binding by Ca2+, CaM wraps around M13, decreasing the distance between the GFP proteins and allowing FRET. In bacteria, the presence of Ca2+ was visualized as a change in emission on violet excitation from 445 nm to 510 nm, and this biosensor reported Ca2+ concentrations from 10–7 to 10–4 M. Independently, and in parallel, Romoser et al. linked blue- and green-emitting GFPs through M13 to detect Ca2+. In the absence of Ca2+, excitation of blue GFP causes the green acceptor to fluoresce at 505 nm as a result of FRET. Binding of Ca2+ and CaM to M13 increases the inter-fluorophore distance, eliminates FRET and causes donor emission at 448 nm. Fluorescence responses to Ca2+ in cells were enhanced by co-expressing CaM with the biosensor. These two Ca2+ biosensors were the first genetically encoded molecular sensors.

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Authors and Affiliations

  1. Associate Editor, Nature Reviews Molecular Cell Biology,

    Katharine H. Wrighton

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  1. Katharine H. Wrighton
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Wrighton, K. Sensing second messengers. Nat Cell Biol 11 (Suppl 1), S20–S21 (2009). https://doi.org/10.1038/ncb1934

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