Skip to main page content
U.S. flag

An official website of the United States government

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Review
. 2005 Dec;8(6):574-81.
doi: 10.1016/j.pbi.2005.09.015. Epub 2005 Sep 26.

Shining light on signaling and metabolic networks by genetically encoded biosensors

Sylvie Lalonde  1 , David W Ehrhardt, Wolf B Frommer
Affiliations
Review

Shining light on signaling and metabolic networks by genetically encoded biosensors

Sylvie Lalonde et al. Curr Opin Plant Biol. 2005 Dec.

Abstract

Fluorescent labels have revolutionized cell biology. Signaling intermediates and metabolites can be measured in real time with subcellular spatial resolution. Most of these sensors are based on fluorescent proteins, and many report fluorescence resonance energy transfer. Because the biosensors are genetically encoded, a toolbox for addressing cell biological questions at the systems level is now available. Fluorescent biosensors are able to determine the localization of proteins and their dynamics, to reveal the cellular and subcellular localization of the respective interactions and activities, and to provide complementary data on the steady state levels of ions, metabolites, and signaling intermediates with high temporal and spatial resolution. They represent the basis for cell-based high-throughput assays that are necessary for a systems perspective on plant cell function.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Fluorescent biosensors for protein–protein interactions. Models for protein–protein interaction biosensors using (a) full-length fluorescent proteins or (b) the split-GFP to reconstitute fluorescent proteins with different spectral properties. Color code: fusion protein (red), ECFP (cyan), EYFP/citrine/Venus (yellow).
Figure 2
Figure 2
Fluorescent biosensors for detecting and localizing protein activities. (a) Sensors for detecting kinase activity, (b) protease activity, and (c) GTPase activation. Color code: ECFP (cyan), EYFP/citrine/Venus (yellow), recognition element (red), MiCy (turquoise) and mKO (orange).
Figure 3
Figure 3
Fluorescent biosensors for metabolites. (a) The calcium sensor cameleon, which consists of the calcium-binding domain of calmodulin fused to myosin light chain. (b) The sensor for cAMP, which uses a guanine nucleotide exchange factor (GEF) as a reporter element. (c) Sensor for the catalytic subunit and regulatory domain of a protein kinase A. (d) The sensor for inositol phosphate/phosphatidylinositol bisphosphate uses the phosphatidylinositol phosphate binding domain of ActA from Listeria. (e) FLIP nanosensors for maltose, glucose, ribose and glutamate that use PBPs. (f) FLIIP nanosensors (for glucose and glutamate) in which CFP was inserted in a permissive site of the periplasmic binding protein. (g) Sensors for pH (CFP–YFP tandem) or for pH and halide (clomeleon) that use a fusion between CFP and YFP. (h) The voltage sensor FlAsH. Color code: ECFP (cyan), EYFP/citrine/Venus (yellow), recognition element (red), MiCy (turquoise) and mKO (orange).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Pertz O, Hahn KM. Designing biosensors for Rho family proteins — deciphering the dynamics of Rho family GTPase activation in living cells. J Cell Sci. 2004;117:1313–1318. - PubMed

    2. This review discusses the different sensors developed for analyzing the dynamics of Rho GTPase activity. The pros and cons of using fluorescent dyes and FPs for successful sensor development are outlined.

    1. Liou J, Kim ML, Heo WD, Jones JT, Myers JW, Ferrell JE, Jr, Meyer T. STIM is a Ca2+ sensor essential for Ca2+-store-depletion-triggered Ca2+ influx. Curr Biol. 2005;15:1235–1241. - PMC - PubMed

    2. This work represents the first systematic analysis of signaling cascades using a high-throughput cell biological assay system in combination with 2300 siRNA targets that are related to putative signaling proteins.

    1. Rizzo MA, Springer GH, Granada B, Piston DW. An improved cyan fluorescent protein variant useful for FRET. Nat Biotechnol. 2004;22:445–449. - PubMed
    1. Nagai T, Ibata K, Park ES, Kubota M, Mikoshiba K, Miyawaki A. A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications. Nat Biotechnol. 2002;20:87–90. - PubMed
    1. Evanko DS, Haydon PG. Elimination of environmental sensitivity in a cameleon FRET-based calcium sensor via replacement of the acceptor with Venus. Cell Calcium. 2005;37:341–348. - PubMed

Publication types

LinkOut - more resources