SpringerProtocols: Abstract: Redox-Sensitive Green Fluorescent Protein: Probes for Dynamic Intracellular Redox Responses. A Review

4. Redox-Sensitive Green Fluorescent Protein: Probes for Dynamic Intracellular Redox Responses. A Review

By: Mark B. Cannon¹, S. James Remington²

Abstract

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The quantification of transient redox events within subcellular compartments, such as those involved in certain signal transduction pathways, requires specific probes with high spatial and temporal resolution. Redox-sensitive variants of the green fluorescent protein (roGFP) have recently been developed that allow the noninvasive monitoring of intracellular thiol-disulfide equilibria. In this chapter, the biophysical properties of these probes are discussed, including recent efforts to enhance their response times. Several recent applications of roGFPs are highlighted, including roGFP expression within Arabidopsis to monitor redox status during root elongation, expression in neurons to measure oxidative stress during ischemia, and targeting of roGFPs to endosomal compartments demonstrating unexpectedly oxidizing potentials within these compartments. Possible future directions for the optimization of roGFPs or new classes of redox-sensitive fluorescent probes are also discussed.

Affiliation(s): (1) Department of Biochemistry and Physical Sciences, BYU-Hawaii, Laie, HI, USA
(2) Institute of Molecular Biology and Department of Physics, University of Oregon, Eugene, OR

Book Title: Redox-Mediated Signal Transduction: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 476 | Year: 2009 | Page Range: 50-64 | DOI: 10.1007/978-1-59745-129-1_4

Subject: Cell Biology

Key Words: Biosensor - disulfide - GFP - midpoint potential - protein engineering - redox

References

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