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This detailed book examines the main methods to study mammalian monoamine oxidases (MAOs), ranging from cell biology to computational chemistry. Beginning with techniques on how to obtain pure samples of MAO A and MAO B, the volume continues by covering assays and techniques used to measure MAO enzymatic activity and perform inhibition studies, methods to address cellular localization and function of MAOs, either in cell lines or in animal models, as well as computational methods applied to rational drug design approaches that are used to develop new MAO inhibitors. Written for 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Monoamine Oxidase: Methods and Protocols serves as a vitalresource for scientists who are interested in studying MAOs and other similar amine oxidase enzymes.
List of contents
Purification of MAO A and of MAO B from Mammalian Tissue Sources.- Purification of Recombinant Eukaryotic MAO A and MAO B Utilizing the Pichia pastoris Expression System.- The Peroxidase-Coupled Assay to Measure MAO Enzymatic Activity.- Measurement of MAO Enzymatic Activity by Spectrophotometric Direct Assays.- Radiochemical Assay of Monoamine Oxidase Activity.- MAO Visible Spectroscopy for Ligand Interactions, Redox Chemistry, and Kinetics of Irreversible Inhibition.- Conventional Receptor Radioligand Binding Techniques Applied to the Study of Monoamine Oxidase.- Assay of MAO Inhibition by Chromatographic Techniques (HPLC/HPLC-MS).- Crystallization of Human Monoamine Oxidase B.- Detecting Monoamine Oxidase A and B Proteins: A Western Blotting Protocol and Some Practical Considerations.- An (Immune)Fluorescence Protocol for Monitoring Monoamine Oxidase A/B Protein Distribution within the Cell.- Expression and Function of MAO A in Cardiac Cells by Means of Adenovirus-Mediated Gene-Transfer.- In Vitro and In Vivo Assays Characterizing MAO A Function in Cancers.- In Vivo Study of Monoamine Oxidases Using Multisite Intracerebral Microdialysis.- Informed Use of 3D-QSAR for the Rational Design of Coumarin Derivatives as Potent and Selective MAO B Inhibitors.- Hansch-Type QSAR Models for the Rational Design of MAO Inhibitors: Basic Principles and Methodology.- Computational Chemistry and Molecular Modeling of Reversible MAO Inhibitors.
