Principles And Methods In Receptor Binding

One of the major advances in the understanding of the mechanism of action of hormones, neurotransmitters and drugs had arisen from the hypothesis that the physiological or pharmacological responses are trig gered by their interaction with specific cell compo nents, termed receptors. However, the presence of receptors has been inferred from indirect data, and only recently has it been possible to study the kinetics of the interaction between drug and receptors directly, through the so called "binding technique. " This NATO-ASI on "Principles and Methods in Receptor Binding" was devoted mainly to the following aspects of the study of receptors: the principles underlying the use of the binding technique; the mathematical models necessary to interpret binding data; the application of binding methods to specific receptors; and, finally, a few selected examples of coupling between receptors and physiological re sponses. In the chapters of this book, special interest is devoted to the analysis of the simplest models for the interaction of receptors with their ligands (either hormones or neurotransmitters or drugs). The graphical techniques used to analyze the data from binding exper iments are extensively discussed, together with the statistics that have to be used in binding analysis. Moreover, the basic concepts to analyze binding data using a personal computer are presented. v vi PREFACE The factors which should be considered when setting up a binding assay (such as choice of ligand and incubation conditions, preparation of tissue, termina tion of incubation) are discussed.

Cell Surface Receptors: A Short Course on Theory and Methods, Second Edition is a primer for the study of cell surface receptors. The simplified discussion of methods and their underlying principles removes the usual intimidation caused by the specialized vocabulary or sophisticated mathematics that characterize many of the primary papers in this field. In this way, the basic concepts become emphasized. This volume is a starting point: a textbook as well as a manual to which the investigator can return for a refresher course, when needed.

This cutting-edge collection of step-by-step experimental protocols demonstrates

This practical reference for medicinal and pharmaceutical chemists combines the theoretical background with modern methods as well as applications from recent lead finding and optimization projects. Divided into two parts on the thermodynamics and kinetics of drug-receptor interaction, the text provides the conceptual and methodological basis for characterizing binding mechanisms for drugs and other bioactive molecules. It covers all currently used methods, from experimental approaches, such as ITC or SPR, right up to the latest computational methods. Case studies of real-life lead or drug development projects are also included so readers can apply the methods learned to their own projects. Finally, the benefits of a thorough binding mode analysis for any drug development project are summarized in an outlook chapter written by the editors.

A broad definition of a receptor is a specialized protein on or in a cell that recognizes and binds a specific ligand to undergo a conformational change, leading to a physiological response or change in cell function. A ligand can be an endogenous neurotransmitter, hormone, paracrine/autocrine factor, or a synthetic drug that may function as an agonist or antagonist. The third edition of Receptor Binding Techniques expands upon the methods and techniques used for studying receptors in silico, in vitro and in vivo. Comprehensive chapters describe how to use online resources for experimental research such as prediction of receptor-ligand interactions and mine the IUPHAR receptor database. Classical techniques of radioligand binding, quantitative autoradiography and their analyses are complemented by the use of immunocytochemistry for the cellular localization of receptor protein and hybridization to detect receptor mRNA. Protocols using fluorescent labeled ligands are described to visualise receptors in living cells, their interaction with beta-arrestin to measure ligand-induced internalisation and green fluorescent protein to study trafficking. Non-radioactive, chemiluminescent cAMP and arrestin assays facilitate the identification of novel ‘biased agonists’. Detailed methods are provided for in vivo imaging of receptors using positron emission tomography (PET). Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Receptor Binding Techniques, Third Edition, aids scientists in continuing to study receptor binding.

New edition of biochemistry textbook which introduces principles and techniques used in undergraduate practical classes.