Real-time imaging of NADPH oxidase activity in living cells using a novel fluorescent protein reporter
| العنوان: | Real-time imaging of NADPH oxidase activity in living cells using a novel fluorescent protein reporter |
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| المؤلفون: | George G. Rodney, Shumin Li, Charles G. Minard, Poulami Basu Thakur, Rituraj Pal |
| المصدر: | PLoS ONE, Vol 8, Iss 5, p e63989 (2013) PLoS ONE |
| بيانات النشر: | Public Library of Science (PLoS), 2013. |
| سنة النشر: | 2013 |
| مصطلحات موضوعية: | Lipopolysaccharides, Anatomy and Physiology, Time Factors, Mouse, Muscle Fibers, Skeletal, lcsh:Medicine, Stimulation, Biosensing Techniques, Green fluorescent protein, RoGFP, Mice, 0302 clinical medicine, Genes, Reporter, Immune Physiology, Molecular Cell Biology, Muscular dystrophy, lcsh:Science, Musculoskeletal System, Image Cytometry, chemistry.chemical_classification, 0303 health sciences, Membrane Glycoproteins, Multidisciplinary, Muscle Biochemistry, Animal Models, respiratory system, Molecular Imaging, 3. Good health, medicine.anatomical_structure, Biochemistry, NADPH Oxidase 2, cardiovascular system, Muscle, Cellular Types, medicine.symptom, Oxidation-Reduction, Research Article, Muscle contraction, circulatory and respiratory physiology, inorganic chemicals, Cell Physiology, Cell Survival, Immune Cells, Green Fluorescent Proteins, In Vitro Techniques, Biology, Fluorescence, 03 medical and health sciences, Model Organisms, Computer Systems, medicine, Animals, Humans, NOx, 030304 developmental biology, Muscle Cells, Reactive oxygen species, lcsh:R, NADPH Oxidases, Skeletal muscle, Macrophage Activation, medicine.disease, Electric Stimulation, chemistry, lcsh:Q, Physiological Processes, Extracellular Space, Reactive Oxygen Species, Cytometry, 030217 neurology & neurosurgery |
| الوصف: | Production of reactive oxygen species (ROS) has been implicated in the pathology of many conditions, including cardiovascular, inflammatory and degenerative diseases, aging, muscular dystrophy, and muscle fatigue. NADPH oxidases (Nox) have recently gained attention as an important source of ROS involved in redox signaling. However, our knowledge of the source of ROS has been limited by the relatively impoverished array of tools available to study them and the limitations of all imaging probes to provide meaningful spatial resolution. By linking redox-sensitive GFP (roGFP) to the Nox organizer protein, p47(phox), we have developed a redox sensitive protein to specifically assess Nox activity (p47-roGFP). Stimulation of murine macrophages with endotoxin resulted in rapid, reversible oxidation of p47-roGFP. In murine skeletal muscle, both passive stretch and repetitive electrical stimulation resulted in oxidation of p47-roGFP. The oxidation of p47-roGFP in both macrophages and skeletal muscle was blocked by a Nox specific peptide inhibitor. Furthermore, expression of p47-roGFP in p47(phox) deficient cells restored Nox activity. As Nox has been linked to pathological redox signaling, our newly developed Nox biosensor will allow for the direct assessment of Nox activity and the development of therapeutic Nox inhibitors. |
| اللغة: | English |
| تدمد: | 1932-6203 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a78ddfe9d3de228148107381af0bb275Test http://europepmc.org/articles/PMC3660327?pdf=renderTest |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....a78ddfe9d3de228148107381af0bb275 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19326203 |
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