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This second edition details new and updated techniques used to study disulfide bonds. Chapters guide readers through how disulfide bonds are classified, techniques used to study functional disulfides, allosteric disulfide bonds, and examples of how labile disulfide bonds are employed for new diagnostics and therapeutics. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Functional Disulphide Bonds: Methods and Protocols, Second Edition serves as a vital guide to this evolving area of study.
List of contents
Classification of Disulfide Bonds.- Structural Analysis of Disulfide Bonds in the RCSB Protein Data Bank Using proteusPy.- Assessing the Evolutionary Conservation of Protein Disulfide Bonds.- Estimate of Numbers of Disulfide-Bonded Protein States.- Using AlphaFold3 for the Interrogation of Protein Disulfide Bonds.- Replacing Cysteine with Selenocysteine: Biochemical Considerations, Computational Modelling, and Protein Engineering Applications.- Quantification of the Redox State of Protein Disulfide Bonds.- Quantification of Site-Specific Disulfide Bond Redox States in Proteins by Parallel Reaction Monitoring-Mass Spectrometry (PRM-MS).- Identification of Target Disulphide Bonds Using Mechanism-Based Kinetic Trapping Combined with Differential Cysteine Labelling.- Quantification of the Redox State of Integrin Disulfide Bonds.- Identification of Protein Cysteine Modifications Using Un-Biased Mass Spectrometry-based Proteomics.- Mapping Protein Disulfide Bonds by Mass Spectrometry.- Determining the Redox Potential of a Protein Disulfide Bond.- Oxidative Protein Folding Using trans-3,4-Dihydroxyselenolane Oxide.- Method for Selection of Antithrombin Disulfide-Bonded Subsets.- Polyclonal Antibody Selection of Partially Disulfide-Bonded Protein States.- Dynamic Force Spectroscopy for Analysis of Multiple Disulfide-Bonded Protein States.- Assays of Thiol Isomerase Activity.- Probing the Mechano-Redox Control of Cell Movement Using Microfluidic Assays.- Flow Cytometry Assessment of Procoagulant Platelets Using a Dithiol-Reactive Probe.- Preparation of a Dithiol-Reactive Probe for PET Imaging of Cell Death.
Summary
This second edition details new and updated techniques used to study disulfide bonds. Chapters guide readers through how disulfide bonds are classified, techniques used to study functional disulfides, allosteric disulfide bonds, and examples of how labile disulfide bonds are employed for new diagnostics and therapeutics. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical,
Functional Disulphide Bonds: Methods and Protocols, Second Edition
serves as a vital guide to this evolving area of study.
