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Titlebook: Oxygen Sensing; Methods and Protocol Emily E. Weinert Book 2023 The Editor(s) (if applicable) and The Author(s), under exclusive license to

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ying OR/MS techniques to EDM.Filar and Haurie are leaders inThe 21st century promises to be an era dominated by international response to c- tain global environmental challenges such as climate change, depleting biodiversity and biocapacity as well as general atmospheric, water and soil pollution pr
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Measurement of O2 Binding by Sensory Hemeproteins,ins. It is critical to measure the affinities of these proteins for ligands like O., CO, and NO, know with confidence when a protein is fully saturated with a specific ligand, and be able to estimate how well a ligand will compete against other ligands for a specific protein. Below we describe how t
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Anaerobic Infrared Spectroelectrochemical Methods for Studying Oxygen-Sensitive [FeFe] Hydrogenasesonments for characterization. Understanding the electrochemical relationships between various active and inactive states of these enzymes is instrumental in uncovering the reaction mechanisms of the complex six-iron active center of [FeFe] hydrogenases called H-cluster. Since states of the H-cluster
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In Vitro Measurement of Gas-Dependent and Redox-Sensitive Diguanylate Cyclase Activity,y messengers cyclic di-GMP. Gas sensing can be difficult to measure due to the high concentration of the oxygen in the atmosphere, particularly in redox-sensitive systems. Here, we describe a method for anaerobic quantification of cyclic di-GMP production which can be used to measure the impact of m
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Spectrophotometric Method for the Quantification and Kinetic Evaluation of In Vitro c-di-GMP Hydrolzation of enzymes under aerobic and anaerobic conditions. The method allows for obtaining all necessary data to calculate . and . from reactions within a single 96-well plate that can be measured using a standard plate reader. The spectrophotometric assay has been used to measure the rates and obtai
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Hydrogen/Deuterium Exchange Mass Spectrometry of Heme-Based Oxygen Sensor Proteins,he isotope exchange of backbone amides. It has virtually no limitations in terms of protein size, flexibility, or reaction conditions and can thus be performed in solution at different pH values and temperatures under controlled redox conditions. Thanks to its coupling with mass spectrometry (MS), i
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Methods for Biophysical Characterization of SznF, a Member of the Heme-Oxygenase-Like Diiron Oxidasve a diferrous cofactor that is coordinated by histidine and carboxylate ligands. Upon exposure to oxygen, the cofactor oxidizes to its diferric state forming a peroxo- adduct, capable of catalyzing a wide range of oxidative chemistries such as desaturation and heteroatom oxidation. Despite their ve
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