Iron Metabolism Methods and Protocols

This detailed volume explores classical and cutting-edge methods optimized and validated to analyze various aspects of iron metabolism, from in vitro to multi-organ level. Opening with a section on basic iron metabolism methods, the book continues with methods applicable to a variety of systems, ranging from bacteria to cultured mammalian cells and tissues, with a focus on cellular heme and iron-sulfur cluster species, as well as mitochondrial iron and its derivatives. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Iron Metabolism: Methods and Protocols serves as an ideal guide for researchers seeking to help in demystifying and boosting iron metabolism-related research.

Iron Homeostatic Regulation in Saccharomyces cerevisiae: Introduction to a Computational Modeling Method.- ICP-MS Analysis of Iron from Biological Samples.- Sample Preparation for X-Ray Fluorescence Microscopy of Iron Distribution in Biological Specimen.- Proximity Ligation Assay for the Analysis of Iron Protein Interactions in the Nucleus.- Fluorescence-Based Proteoliposome Methods to Monitor Redox-Active Transition Metal Transmembrane Translocation by Metal Transporters.- Metal Uptake by Mitochondrial Carrier Family Proteins Using Lactococcus lactis.- Methods for Cultivation of Bacteroides thetaiotaomicron and Analysis of Heme Metabolism by Mass Spectrometry and Spectroscopic Approaches.- Heme o Isolation and Analysis Using Heme o Synthase Heterologously Expressed in Escherichia coli.- Fluorometric Methods to Measure Bioavailable and Total Heme.- Recombinant Biogenesis and Analysis of Cytochrome c Species.- Ultra Performance Liquid Chromatography (UPLC) Analysis of Heme Biosynthesis Intermediates.- Radiolabeling Heme and Porphyrin with 14C-Glycine or 14C -Aminolevulinic Acid.- Heterologous Expression and Purification of Eukaryotic ALA Synthase from E. coli.- Direct Spectroscopic Ferrochelatase Assay.- Reverse Thiol Trapping Approach to Assess the Thiol Status of Metal-Binding Mitochondrial Proteins.- A Chemoproteomic Approach to Monitor Native Iron-Sulfur Cluster Binding.