Angiogenin (ANG) is a vertebrate-specific ribonuclease possessing both ribonucleolytic, neuroprotective, and angiogenic activity. ANG binds to and is inhibited by the ribonuclease inhibitor protein (RNH1), a horseshoe-shaped leucine-rich repeat protein with marked oxidation sensitivity. Both ANG and RNH1 respond to various types of cell stress, including oxidation, starvation, and hypoxia. The goal of this work is to characterize the real-time changes of ANG distribution in cells under starvation and oxidative stress conditions. We have developed fluorescence microscopy techniques in live HeLa cells using an educational Warwick Open Source Microscope (eduWOSM), an inexpensive 3D-printed microscope with single-molecule detection capabilities. Our current work is aimed at characterizing the internalization and localization of fluorescently-labeled ANG in HeLa cells. Co-localization of RNH1 will be detected by Western Blots in fractionated HeLa cells and confirmed by co-immunoprecipitation of Flag-ANG:RNH1 complexes. The Lawrence University Chandler Senior Experience Fund, supplemented by my Student Chapter Travel Award, is the source that funded my research.
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Lagman et al. (2024) studied this question.
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