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

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.

[1]  Jin Zhang,et al.  Chapter 2: Molecular sensors based on fluorescence resonance energy transfer to visualize cellular dynamics. , 2008, Methods in cell biology.

[2]  Lakshmi Krishnamoorthy,et al.  A Cellular FRET-Based Sensor for β-O-GlcNAc, A Dynamic Carbohydrate Modification Involved in Signaling , 2006 .

[3]  M. Davidson,et al.  Advances in fluorescent protein technology , 2011, Journal of Cell Science.

[4]  R. Lefkowitz,et al.  Trafficking patterns of beta-arrestin and G protein-coupled receptors determined by the kinetics of beta-arrestin deubiquitination. , 2003, The Journal of biological chemistry.

[5]  Jin Zhang,et al.  FRET-based biosensors for protein kinases: illuminating the kinome. , 2007, Molecular bioSystems.

[6]  A. Sickmann,et al.  Modificomics: posttranslational modifications beyond protein phosphorylation and glycosylation. , 2007, Biomolecular engineering.

[7]  Juergen Kast,et al.  Current approaches for global post-translational modification discovery and mass spectrometric analysis. , 2008, Analytica chimica acta.

[8]  K. Svoboda,et al.  A genetically encoded fluorescent sensor of ERK activity , 2008, Proceedings of the National Academy of Sciences.

[9]  R. Lefkowitz,et al.  Trafficking Patterns of β-Arrestin and G Protein-coupled Receptors Determined by the Kinetics of β-Arrestin Deubiquitination* , 2003, The Journal of Biological Chemistry.

[10]  Kotaro Oka,et al.  Simultaneous Live Cell Imaging Using Dual FRET Sensors with a Single Excitation Light , 2009, PloS one.

[11]  A. Ting,et al.  Genetically encoded fluorescent reporters of histone methylation in living cells. , 2004, Journal of the American Chemical Society.

[12]  Michel Bouvier,et al.  Real-time monitoring of ubiquitination in living cells by BRET , 2004, Nature Methods.

[13]  A. Miyawaki,et al.  Expanded dynamic range of fluorescent indicators for Ca(2+) by circularly permuted yellow fluorescent proteins. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Robert E Campbell,et al.  Genetically encoded FRET-based biosensors for multiparameter fluorescence imaging. , 2009, Current opinion in biotechnology.

[15]  Xiang-Jiao Yang,et al.  Multisite protein modification and intramolecular signaling , 2005, Oncogene.

[16]  R. Lefkowitz,et al.  Seven transmembrane receptors: something old, something new , 2007, Acta physiologica.

[17]  M. Hochstrasser,et al.  Modification of proteins by ubiquitin and ubiquitin-like proteins. , 2006, Annual review of cell and developmental biology.

[18]  G. Hart,et al.  O-GlcNAcylation regulates phosphorylation of tau: a mechanism involved in Alzheimer's disease. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[19]  Shu Chien,et al.  Determination of hierarchical relationship of Src and Rac at subcellular locations with FRET biosensors , 2008, Proceedings of the National Academy of Sciences.

[20]  Necmettin Yildirim,et al.  β2-Adrenergic Receptor Signaling and Desensitization Elucidated by Quantitative Modeling of Real Time cAMP Dynamics* , 2008, Journal of Biological Chemistry.

[21]  T. Kerppola,et al.  Ubiquitin-mediated fluorescence complementation reveals that Jun ubiquitinated by Itch/AIP4 is localized to lysosomes. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Y. Kanno,et al.  Selective recognition of acetylated histones by bromodomain proteins visualized in living cells. , 2004, Molecular cell.

[23]  B. Kobilka,et al.  Myocyte Adrenoceptor Signaling Pathways , 2003, Science.

[24]  T. Pawson,et al.  Reading protein modifications with interaction domains , 2006, Nature Reviews Molecular Cell Biology.

[25]  Jun O. Liu,et al.  Reading dynamic kinase activity in living cells for high-throughput screening. , 2006, ACS chemical biology.

[26]  F Wold,et al.  In vivo chemical modification of proteins (post-translational modification). , 1981, Annual review of biochemistry.

[27]  Roger Y Tsien,et al.  Insulin disrupts beta-adrenergic signalling to protein kinase A in adipocytes. , 2005, Nature.

[28]  Christopher T. Walsh,et al.  Protein Posttranslational Modifications: The Chemistry of Proteome Diversifications , 2006 .

[29]  David L. Brautigan,et al.  Midzone activation of aurora B in anaphase produces an intracellular phosphorylation gradient , 2008, Nature.

[30]  H. Funabiki,et al.  Correcting aberrant kinetochore microtubule attachments: an Aurora B-centric view. , 2009, Current opinion in cell biology.

[31]  J. Lakowicz.  Principles of fluorescence spectroscopy , 1983 .

[32]  G. Hart,et al.  Elevated nucleocytoplasmic glycosylation by O-GlcNAc results in insulin resistance associated with defects in Akt activation in 3T3-L1 adipocytes , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Jin Zhang,et al.  Dynamic Protein Kinase A Activities Induced by β-Adrenoceptors Dictate Signaling Propagation for Substrate Phosphorylation and Myocyte Contraction , 2009, Circulation research.

[34]  Amy E Palmer,et al.  Fluorescent biosensors of protein function. , 2008, Current opinion in chemical biology.

[35]  Jin Zhang,et al.  Spatiotemporal analysis of differential Akt regulation in plasma membrane microdomains. , 2008, Molecular biology of the cell.

[36]  Roger Y. Tsien,et al.  A genetically encoded fluorescent reporter reveals oscillatory phosphorylation by protein kinase C , 2003, The Journal of cell biology.

[37]  Roger Y. Tsien,et al.  Insulin disrupts β-adrenergic signalling to protein kinase A in adipocytes , 2005, Nature.

[38]  R. Newman,et al.  Visualization of phosphatase activity in living cells with a FRET-based calcineurin activity sensor. , 2008, Molecular bioSystems.

[39]  B. Kobilka,et al.  Phosphodiesterase 4D is required for beta2 adrenoceptor subtype-specific signaling in cardiac myocytes. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[40]  A. Ting,et al.  A genetically encoded fluorescent reporter of histone phosphorylation in living cells. , 2004, Angewandte Chemie.