Light-Activated Regulation of Cofilin Dynamics Using a Photocaged Hydrogen Peroxide Generator

Miller, Evan W.; Taulet, Nicolas; Onak, Carl S.; New, Elizabeth J.; Lanselle, Julie K.; Smelick, Gillian S.; Chang, Christopher J.

doi:10.1021/ja107783j

Cited by 33 publications

(31 citation statements)

References 39 publications

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“…General oxidative stress photosensitizers that target nuclei and mitochondria were created using organelle-specific peptide delivery systems 91 . In parallel, our laboratory has developed a caged small-molecule H 2 O 2 generator that rapidly produces H 2 O 2 on demand upon cleavage of a photolabile protecting group 92 . We used this new reagent to produce H 2 O 2 in living cells by light activation and trigger downstream redox regulation of cofilin, leading to actin polymerization and cell migration.…”

Section: Chemical Tools For Studying Ros Biologymentioning

confidence: 99%

Chemistry and biology of reactive oxygen species in signaling or stress responses

2011

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Reactive oxygen species (ROS) are a family of molecules that are continuously generated, transformed and consumed in all living organisms as a consequence of aerobic life. The traditional view of these reactive oxygen metabolites is one of oxidative stress and damage that leads to decline of tissue and organ systems in aging and disease. However, emerging data show that ROS produced in certain situations can also contribute to physiology and increased fitness. This Perspective provides a focused discussion on what factors lead ROS molecules to become signal and/or stress agents, highlighting how increasing knowledge of the underlying chemistry of ROS can lead to advances in understanding their disparate contributions to biology. An important facet of this emerging area at the chemistry-biology interface is the development of new tools to study these small molecules and their reactivity in complex biological systems.

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Section: Chemical Tools For Studying Ros Biologymentioning

confidence: 99%

Chemistry and biology of reactive oxygen species in signaling or stress responses

2011

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“…Miyata and co‐workers reported the formation of NO by the two‐photon excitation of fluorescein linked to 2,6‐dimethylnitrobenzene 102. Chang and co‐workers caged 1,2,4‐trihydroxybenzene, which reduces molecular oxygen to superoxide and leads to the formation of H 2 O 2 103. H 2 O 2 activates cofilin, an actin depolymerization factor at the actin rods in HeLa cells.…”

Section: Irreversible Photo(de)activation (Uncaging)mentioning

confidence: 99%

Light‐Controlled Tools

et al. 2012

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Spatial and temporal control over chemical and biological processes plays a key role in life, where the whole is often much more than the sum of its parts. Quite trivially, the molecules of a cell do not form a living system if they are only arranged in a random fashion. If we want to understand these relationships and especially the problems arising from malfunction, tools are necessary that allow us to design sophisticated experiments that address these questions. Highly valuable in this respect are external triggers that enable us to precisely determine where, when, and to what extent a process is started or stopped. Light is an ideal external trigger: It is highly selective and if applied correctly also harmless. It can be generated and manipulated with well-established techniques, and many ways exist to apply light to living systems--from cells to higher organisms. This Review will focus on developments over the last six years and includes discussions on the underlying technologies as well as their applications.

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“…Thus, microscopic analysis is the preferred measurement technique because it leaves the samples unperturbed. Most prior studies that included microscopic image analysis of cells expressing peroxide sensors used small sample sizes (~10 cells) (19,20,(29)(30)(31)(32). To facilitate analysis of larger sample sizes, we automated the image analysis process, using a combination of ImageJ (National Institutes of Health, Bethesda, MD) and CellProfiler software (Broad Institute).…”

Section: Analysis Of Fluorescence Imagesmentioning

confidence: 99%

Interpreting Heterogeneity in Response of Cells Expressing a Fluorescent Hydrogen Peroxide Biosensor

Huang

Ali

Stein

et al. 2015

Biophysical Journal

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Fluorescent, genetically encoded sensors of hydrogen peroxide have enabled visualization of perturbations to the intracellular level of this signaling molecule with subcellular and temporal resolution. Ratiometric sensors hold the additional promise of meaningful quantification of intracellular hydrogen peroxide levels as a function of time, a longstanding goal in the field of redox signaling. To date, studies that have connected the magnitudes of observed ratios with peroxide concentrations have either examined suspensions of cells or small numbers of adherent cells (∼10). In this work, we examined the response of all cells in several microscopic fields of view to an identical perturbation and observed a striking degree of heterogeneity of fluorescence ratios from individual cells. The expression level of the probe and phase within the cell cycle were each examined as potential contributors to the observed heterogeneity. Higher ratiometric responses correlated with greater expression levels of the probe and phase in the cell cycle were also shown to influence the magnitude of response. To aid in the interpretation of experimental observations, we incorporated the reaction of the reduced probe with peroxide and the reactions of the oxidized probe with glutathione and glutaredoxin into a larger kinetic model of peroxide metabolism. The predictions of the kinetic model suggest possible explanations for the experimental observations. This work highlights the importance of a systems-level approach to understanding the output of genetically encoded sensors that function via redox reactions involving thiol and disulfide groups.

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Light-Activated Regulation of Cofilin Dynamics Using a Photocaged Hydrogen Peroxide Generator

Cited by 33 publications

References 39 publications

Chemistry and biology of reactive oxygen species in signaling or stress responses

Chemistry and biology of reactive oxygen species in signaling or stress responses

Light‐Controlled Tools

Interpreting Heterogeneity in Response of Cells Expressing a Fluorescent Hydrogen Peroxide Biosensor

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