Year: 2018
Type: article
Abstract: AIMS: Quantitative imaging of glutathione (GSH) with high spatial and temporal resolution is essential for studying the roles of GSH in redox biology. To study the long-standing question of compartmentalization of GSH, especially its distribution between the nucleus and cytosol, an organelle-targeted quantitative probe is needed. RESULTS: We developed a reversible reaction-based ratiometric fluorescent probe-HaloRT-that can quantitatively measure GSH dynamics with subcellular resolution in real time. Using HaloRT, we quantitatively measured the GSH concentrations in the nucleus and cytosol of HeLa cells and primary hepatocytes under different treatment conditions and found no appreciable concentration gradients between these two organelles. Innovation and Conclusion: We developed the first reversible ratiometric GSH probe that can be universally targeted to any organelle of interest. Taking advantage of this new tool, we provided definitive evidence showing that GSH concentrations are not significantly different between the nucleus and cytosol, challenging the view of nuclear compartmentalization of GSH.
Source: Antioxidants and Redox Signaling
Language: English
Topic: Sulfur Compounds in Biology
Subfield: Biochemistry
Domain: Life Sciences
Open Access status: green
Funders: Welch Foundation, Cancer Prevention and Research Institute of Texas, National Institutes of Health, Intellectual and Developmental Disabilities Research Center, National Institute on Aging, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Allergy and Infectious Diseases, National Institute of Diabetes and Digestive and Kidney Diseases, National Institute of Biomedical Imaging and Bioengineering, National Institute of Child Health and Human Development, National Center for Research Resources, National Center for Complementary and Integrative Health, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: G1194650153, G1289049516, Delineate the regulatory network of lipid metabolism via SRS imaging-sorting, G2053725305, G2087349970, G2137056809, G2174142799, BD Biosciences Special Order LSRII, G2421591455, G2711403535, Engineering Antibody Drug Conjugates To Target P53-Defective Triple Negative Breast Cancer, G4444814008, G4641643908, G481577519, G530820192, G5890830462, G5905248492, G617376388, G641713321, G7618450163, Blocking TNFR1 increases tumor radiosensitivity and ameliorates radiation-induced cognitive impairments, G863484009, G8733582291, G8827130515, G8919310273, G9000009718, G9000066955, G9000177741, G9000198215, G9000240210, G9000241107, G9000286996, G9000296792, G9000300963, Baylor College of Medicine Cancer Center - Cancer Center Support Grant, Advanced Computational Approaches for NMR Data-mining, G964973908, Prebiotic Regulation of Longevity, G99424851
OpenAlex ID: W2950627161