First Application of Terephthalate as a Fluorescent Probe for Hydroxyl Radicals in Thylakoid Membranes
The Method Of Hydroxyl Radical Detection Via The Conversion Of Terephthalate (Tpa) Into The Strongly Fluorescent Hydroxyterephthalate Has Been Used For A Long Time In Radiation Research. However, Attempts To Apply It In Biological Research Appeared Only Recently. Possibilities And Limitations Of The Tpa Method Were Investigated In Order To Adapt It For Chlorophyll-Containing Samples. Using Model Chemical Sources Of Various Reactive Oxygen Species We Confirmed That Tpa Very Sensitively Detects Hydroxyl Radicals, But Is Not Reactive To Either Hydrogen Peroxide Or Superoxide. Moreover, Using Rose Bengal As A Photosensitiser, We Showed That Tpa Is Also Insensitive To Singlet Oxygen. Here We Suggest The Optimal Conditions For Using The Method In Thylakoid Preparations And These Conditions Were Applied To Detect The Production Of Hydroxyl Radicals Induced By 365 Nm Uv Radiation. The High Selectivity Of Terephthalate Hydroxylation Towards Hydroxyl Radicals, The Relatively Low ToxiCity, Light-Insensitivity And Chemical Stability Of Both Probe And Its Hydroxylated Form Make This Simple Method Highly Promising For Application In In Vitro Studies.
KeywordsOxidative stress hydroxyl radicals UV-A terephthalate thylakoids
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