Front Cover; Fe-S Cluster Enzymes Part A; Copyright; Contents; Contributors; Preface; Chapter One: Genetic, Biochemical, and Biophysical Methods for Studying Fe-S Proteins and Their Assembly; 1. Fe-S Cluster Biology; 2. Genetic Approaches and Strategies; 2.1. Genetic Selection and/or Screens Based on Fe-S Biogenesis Defects; 2.2. The Use of Conditional Alleles; 2.3. Genetic Approaches to Investigate Functional Redundancy; 2.4. Suppressor-Based Strategies; 2.5. trans-Dominant Mutations; 2.6. E. coli Strains of Particular Interest for Fe-S Cluster Biogenesis Studies.

2.6.1. The eSUF and eISC Strains2.6.2. The Hpx Strain; 2.6.3. The Iron-Minus Strain; 2.7. Construction of Frataxin Models; 3. Biochemistry of Fe-S Clusters; 3.1. Purification of Fe-S Cluster-Containing Proteins; 3.2. Characterization of Fe-S Proteins; 4. Spectroscopy of Fe-S Proteins; 4.1. UV-Visible Absorption Spectroscopy; 4.2. Mössbauer Spectroscopy; 4.3. Electronic Paramagnetic Resonance Spectroscopy; 4.4. Hyperfine Sublevel Correlation Spectroscopy; 4.5. Raman Resonance Spectroscopy; 4.6. Circular Dichroism; 4.7. Nuclear Magnetic Resonance Spectroscopy; Acknowledgments; References.

Chapter Two: De Novo Design of Iron-Sulfur Proteins1. Introduction; 2. [4Fe-4S]; 3. [3Fe-4S] Clusters; 4. [2Fe-2S] Clusters; 5. [Fe8S7] Clusters; 6. Protocols; 6.1. Diiron Hexacarbonyl Cluster Incorporation Into Peptide (Roy et al., 2012); 6.2. [4Fe-4S] Cluster Incorporation Into Peptide; 6.3. EPR [4Fe-4S] Sample Preparation; 6.4. Hydrogen Production With Diiron Hexacarbonyl; 6.5. Electrochemical Measurements; References; Chapter Three: In Vitro Studies of Cellular Iron-Sulfur Cluster Biosynthesis, Trafficking, and Transport; 1. Introduction.

2. Preparation of Fe-S Clusters and Kinetic Determination of Fe-S Cluster Transfer Reactions2.1. Na6[Fe2S2](GS)4 Synthesis; 2.1.1. Equipment; 2.1.2. Buffers and Reagents; 2.1.3. Procedure; 2.1.4. Notes; 2.2. Cluster Uptake and Protein-Protein Transfer Reactions; 2.2.1. Equipment; 2.2.2. Buffers and Reagents; 2.2.3. Procedure; 2.2.4. Notes; 2.3. Glutathione Extraction; 2.3.1. Equipment; 2.3.2. Buffers and Reagents; 2.3.3. Procedure; 2.3.4. Notes; 3. Fe-S Cluster Stimulation and Transport in Liposome Assays; 3.1. ATPase Stimulation Assays; 3.1.1. Equipment; 3.1.2. Buffers and Reagents.

3.1.3. Procedure3.1.4. Notes; 3.2. Liposome Synthesis and Protein Incorporation; 3.2.1. Equipment; 3.2.2. Buffers and Reagents; 3.2.3. Procedure; 3.2.4. Notes; 3.3. Cluster Transfer Monitored by Tiron Assay and Flow Cytometry; 3.3.1. Equipment; 3.3.2. Buffers and Reagents; 3.3.3. Procedure; 3.3.4. Notes; 4. Summary and Conclusions; References; Chapter Four: Combined Biochemical, Biophysical, and Cellular Methods to Study Fe-S Cluster Transfer and Cytosolic Aconit ... ; 1. Introduction; 2. In Vitro Fe-S Cluster Transfer From mNT to a Model Recipient; 2.1. Purifications of mNT and FDX.