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This volume provides researchers with protocols that help them investigate known or putative O2 sensing proteins and pathways. The chapters in this book discuss techniques ranging from anaerobic redox midpoint measurement to approaches to control expression of globin genes, which provide detailed methods for researchers interested in expanding their knowledge of O2 sensing systems. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and comprehensive, Oxygen Sensing: Methods and Protocols is a valuable resource to a wide audience, ranging from microbiologists and cell biologists, to protein biochemists.
List of contents
Integrating UV-Vis Spectroscopy and Oxygen Optode for Accurate Determination of Oxygen Affinity of Proteins.- Measurement of O2 Binding by Sensory Hemeproteins.- Resonance Raman Characterization of O2-Binding Heme Proteins.- Anaerobic Infra-Red Spectroelectrochemical Methods for Studying Oxygen-Sensitive [FeFe] Hydrogenases.- Monitoring the Kinase Activity of Heme-Based Oxygen Sensors and Its Dependence on O2 and Other Ligands using Phos-tag Electrophoresis.- In Vitro Measurement of Gas-Dependent and Redox-Sensitive Diguanylate Cyclase Activity.- Spectrophotometric Method for the Quantification and Kinetic Evaluation of In Vitro c-di-GMP Hydrolysis in the Presence and Absence of Oxygen.- Hydrogen/Deuterium Exchange Mass Spectrometry of Heme-Based Oxygen Sensor Proteins.- Methods for Biophysical Characterization of SznF, A Member of the Heme-Oxygenase-Like Diiron Oxidase/Oxygenase Superfamily.- Analyzing Iron and Oxygen-Regulated ProteinComplex Formation using Proteomic Mass Spectrometry.- CRISPR Activator Approaches to Study Endogenous Androglobin Gene Regulation.- Assays to Study Hypoxia-Inducible Factor Prolyl Hydroxylase Domain 2 (PHD2), A Key Human Oxygen Sensing Protein.- Kinetic Measurement Investigating the Oxygen-Sensing Properties of the Plant Cysteine Oxidases.- Methods for Culturing Anaerobic Microorganisms.
